GENETICS CORE FACILITIES CATALOG Cell Center Service Facility DNA Sequencing Facility Genetic Diagnostic Laboratory Transgenic & Chimeric Mouse Facility and the Cell Center Stockroom 2004 UNIVERSITY OF PENNSYLVANIA DEPARTMENT OF GENETICS Haig H. Kazazian, Jr., M.D. Chairman, Department of Genetics Director, DNA Sequencing Facility 475 Clinical Research Building 215/898-3582 David Gasser, Ph.D. Professor, Department of Genetics Director, Cell Center Service Facility 575 Clinical Research Building 215/898-5175 gasserd@mail.med.upenn.edu Tapan Ganguly, Ph.D. Technical Director, DNA Sequencing Facility and Cell Center Service Facility D6 Richards Building 215/573-7238 gangulyt@mail.med.upenn.edu Barbara Nevin Manager, Cell Center Stockroom D1 Richards Building 215/898-0281 nevin@mail.med.upenn.edu - Please note that Barbara Nevin should be your first contact for issues regarding the Cell Center Stockroom. - We appreciate your suggestions regarding ways in which we can increase the effectiveness of the Cell Center’s services. Your input, addressed to either us or to your department or Center representative on the Advisory Committee, would be helpful and appreciated. Thank you for your continued support. 2004 GENETICS CORE FACILITIES CATALOG TABLE OF CONTENTS DNA Sequencing Facility ............................................................................... 4 Description of Services ...................................................................................................... Charges for Services .......................................................................................................... Recommended Procedures for Template Preparation ....................................................... Network Connections ........................................................................................................ Guide for Oligonucleotide Synthesis ................................................................................ 5 7 8 12 13 Cell Center Services Facility.......................................................................... 17 Cell Center Services and Shared Resources ...................................................................... 17 Summary of Charges for Services ..................................................................................... 21 Genetic Diagnostic Laboratory ..................................................................... 22 Transgenic & Chimeric Animal Facility....................................................... 31 Cell Center Stockroom .................................................................................. 48 Opening an Account ........................................................................................................... 48 How to Order ...................................................................................................................... 49 Order Pickup and Delivery Information ............................................................................. 50 Oligonucleotide Synthesis .................................................................................................. 51 Primers & Probes ................................................................................................................ 54 Stocked Products - Product Class Listings .................................................. 57 Chemicals and Reagents • (CH).......................................................................................... 58 Competent Cells • (CE) ...................................................................................................... 57 Film and Membranes • (FM) .............................................................................................. 63 Immunological Reagents • (IM) ......................................................................................... 64 Miscellaneous Products • (MS) .......................................................................................... 68 Molecular Biology Kits • (KT) ........................................................................................... 65 Molecular Biology Products • (MB) .................................................................................. 66 Non-radioactive Kits and Reagents • (NR). ....................................................................... 69 Other Enzymes • (OE) ........................................................................................................ 69 PCR Reagents and Nucleotides • (PC) ............................................................................... 71 Precast Gels • (PG) ............................................................................................................ 74 Protein & Molecular Weight Markers • (MR) .................................................................... 67 Purification Kits and Columns • (CO)................................................................................ 62 Restriction Enzymes • (RE) ............................................................................................... 74 Sera • (SM) ........................................................................................................................ 78 Tissue Culture Media and Reagents • (TC) ....................................................................... 78 Stocked Products - Listed by Vendor ........................................................... 80 Amersham Biosciences • (AMR) .................…………….................................................. 80 Applied Biosystems • (PEC) ............................................................................................ 101 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 2 Bio-Rad Laboratories • (RAD) ........................................................................................ 106 Biowhittaker Molecular Applications • (BMA) ................................................................ 85 Cambrex Biosciences • (FMC) ......................................................................................... 86 Cell Center Services • (CCS) ............................................................................................. 86 Clontech Laboratories • (CLN) .......................................................................................... 87 Collaborative Biomedical Products • (COL) ..................................................................... 87 Denville Scientific • (DEN) ............................................................................................... 88 Difco Laboratories • (DIF) ................................................................................................. 89 Enzo Life Sceinces • (ENZ) ............................................................................................... 89 Epicentre Technologies • (EPI) ......................................................................................... 89 Fisher Scientific • (FSH) .................................................................................................... 89 Fuji Film • (FUJ) ................................................................................................................ 91 Gemini Bioproducts • (GEM) ............................................................................................ 91 Hyclone Laboratories • (HYC) .......................................................................................... 91 IGEN International (Fisher Scientific) • (IGN) ................................................................. 91 Invitrogen (Life Techologies) • (INV) ............................................................................... 91 Kodak • (KOD) .....................................................................................…......................... 97 Marligen Biosciences • (MLG)........................................................................................... 97 Mediatech (Fisher Scientific) • (MED) .............................................................................. 97 Micron Separations Inc. (Fisher Scientific) • (MSI) .......................................................... 98 Millipore • (MIL) ................................................................................................................ 98 National Diagnostics • (ND) ............................................................................................... 99 New England Biolabs • (NEB) ........................................................................................... 99 Perkin Elmer (New England Nuclear) • (NEN) ................................................................ 101 Polaroid Film • (POL) ....................................................................................................... 102 Promega • (PRO)....................................................................................…........................ 103 Q-Biogene • (DEN) ............................................................................................................ 88 Qiagen • (QIA) .................................................................................................................. 105 Rainin Instruments • (RAI) ............................................................................................... 107 Roche Diagnostics • (BMB) .........................………………………………………......... 82 Sigma-Aldrich • (SIG) ...................................................................................................... 107 Stratagene • (STR) ..................................................................................…...................... 108 TKR Bioproducts • (DEN) ................................................................................................ 88 USB Corporation • (USB) ............................................................................................... 109 Index of Stocked Products .......................................................................... 109 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 3 DNA SEQUENCING FACILITY Haig H. Kazazian, Jr., M.D., Director 215/898-3582 kazazian@mail.med.upenn.edu Tapan Ganguly, Ph.D., Technical Director 215/573-7238 dnaseq@mail.med.upenn.edu, gangulyt@mail.med.upenn.edu Peiqui Chen, Research Specialist 215/573-7407 pchen2@mail.med.upenn.edu Latonia Clayton, Research Technician 215/573-7407 latonia@mail.med.upenn.edu Deborah McKinney, Administrative Assistant 215/573-7407 dlmckinn@mail.med.upenn.edu Elias Papaioannou, Research Specialist 215/573-7407 eliasp2@mail.med.upenn.edu Lanping Zhang, Research Specialist 215/573-7407 lpzhang@mail.med.upenn.edu David Zhu, Research Specialist 215/573-7407 zhu2@mail.med.upenn.edu Location: B1 Richards Building Mailing Address: B1 Richards Building 3700 Hamilton Walk Philadelphia, PA 19104-6085 Telephone: 215/573-7407 Fax: 215/573-9327 Internet: http://www.med.upenn.edu/dnaseq FTP: dnaseq.med.upenn.edu Fast turnaround • Consultation and technical support • Automated cycle sequencing from PCR products to BACs • Sequencing large fragments & project management • In-house primer & oligonucleotide synthesis • Degenerate, inosine, phosphorothioate, biotinylated, amino linked, and fluorescent labeled oligos • Template preparation and purification • GeneScanning for VNTRs • Identification of heterozygosity • Assembly and analysis of sequences, ORF analysis • Genbank homology searches • Submission of new sequences to Genbank University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 4 DNA Sequencing Facility Description of Services _ Please fill out a “Request for Services” form appropriate to each service _ Please acknowledge services performed by the facility in your publications New Service We are now offering Site-directed Mutagenesis as a service using Stratagene’s Quick Change kit. This includes design and synthesis of the mutagenic primers, mutagenesis and verification of the mutation by sequencing. Synthesis and PAGE purification of the primers are done by LTI (BRL). Please contact us for details. Also, we are now in the process of setting up Transgenomic’s DHPLC system for mutation detection, DNA fragment separation, and oligonucleotide purification. Automated Cycle Sequencing The facility uses ABI 377 and 373A Stretch sequencers, with Taq FS Big Dye™ Terminator or Dye Primer chemistry. Please consult our recommendations for template preparation - to avoid problems. Standard primers (listed below) are provided by the facility at no charge. Custom primers should be provided by the user, or can be synthesized at the facility. Facility users can choose between short (~500 nucl accurate read) or long (~600 nucl accurate read) gels. Ready-to-load samples are analyzed at a lower charge. Turn around time is generally 2 working days for sequencing. Color printouts of analyzed chromatograms are provided for all successful reactions. Sequencing results can also be obtained on floppy or Zip disc, or over PennNet, or by FTP for further examination and manipulation by the user. Facility staff will consult with users whenever there are problems, and will make suggestions to improve sequencing results. A good template yields 600 nucleotides of reliable sequence (<2% error and ambiguity) in a long read run. Templates or template-primer mixes for each reaction must be in a separate 1.5 ml tube. Our ability to provide fast turnaround and reliable sequencing service depends on you! Please provide the correct amounts and volumes of templates and primers (as indicated on the Sequencing Request Form), and make sure all the requested information on the form is completed. The following standard primers provided by the Sequencing Facility at no charge are listed in Table 1. Template Preparation and Purification Template DNA can be prepared and/or purified by the facility staff. Low cost plasmid minipreps processed on the QIAGEN BioRobot are offered with or without sequencing. Larger scale purification is also available. Lambda phage cosmid, PI and BAC preps of comparable scales are also performed. PCR reactions and purification using commercial columns, with or without gel analysis, is also offered by the facility. The facility also offers a reliable treatment of marginal or bad templates to make them suitable for automated sequencing. This involves RNase treatment followed by column purification or phenol-chloroform extraction. This treatment improves the quality of the template, producing more reliable sequence and longer reads. For primer University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 5 walking projects we recommend that a substantial amount of DNA (nominally 4-5 µg per kb for each direction) be treated this way and be used for the whole project. Templates prepared or purified by the facility staff are guaranteed to produce good sequence with standard, reliable primers. The facility also provides free training in template preparation and troubleshooting at the user’s request. Oligonucleotide Synthesis Two ABI 394 four-column synthesizers are used for synthesizing primers for sequencing projects and oligos requested by researchers. After synthesis, oligonucleotides are unprotected, yield is measured by OD260, and the oligo is desalted (if necessary) and dried. Affinity column OPC purification of full-length oligos is available if requested. Each column can be loaded with 5-15 OD260 of oligo, and will yield 3-8 OD260 after purification. DegenTABLE 1: Standard Primers Available at No Charge Primer Sequence Bases M.W. TmoC M13/pUC Forward 5’ CCCAG TCACG ACGTT GTAAA ACG 3’ 23 6978 69.2 M13/pUC Reverse 5’ AGCGG ATAAC AATTT CACAC AGG 3’ 23 7066 65.8 Sp6 5’ ATTTA GGTGA CACTA TAGAA 3’ 20 6164 47.6 T3 promoter/primer 5’ ATTAA CCCTC ACTAA AGGGA 3’ 20 6094 56.7 T7 promoter/primer 5’ TAATA CGACT CACTA TAGGG 3’ 20 6125 50.8 T7 terminator 5’ GCTAG TTATT GCTCA GCGG 3’ 19 5830 60.1 gt 10 Forward 5’ CCTTT TGAGC AAGTT CAGCC TGG 3’ 23 7025 69.7 gt 10 Reverse 5’ GGTGG CTTAT GAGTA TTTCT TCC 3’ 23 7046 61.5 gt 11 Forward 5’ ATTGG TGGCG ACGAC TCCTG GAG 3’ 23 7112 74.2 gt 11 Reverse 5’ CAGAC CAACT GGTAA TGGTA GC 3’ 22 6763 62.2 DNA SEQUENCING FACILITY 7 erate, inosine containing, phosphorothioate, biotinylated, 5’or 3’ phosphorylated oligos, and oligos with terminal amino groups or fluorescent tags can be synthesized. Turnaround time is typically 2-3 days. Large Sequencing Projects The facility will sequence through DNA clones by primer walking and/or subcloning. Please fill out a Request for Sequencing Project Management form when submitting the sample, indicating if you wish to select primer sequences yourself or delegate the responsibility to facility staff. Charges for primer walks are calculated according to the number of lanes sequenced and primers synthesized. A clean-up treatment of sufficient template for the whole project (nominally 4-5 µg per kb for each direction) is recommended to produce quality sequence and maximize step size. After good quality template is obtained, each step (~450 nucl.) requires about a week, including primer TABLE 2: Charges for Services Cancer Center Other & Genetics Members Investigators DNA Sequencing Long Run DNA Sequencing (reaction, purification, electrophoresis) .............................. $17.50/sample $19.50/sample DNA Sequencing Reaction Done (purification, electrophoresis) .................... $12.50/sample $14/sample DNA Sequencing Ready to Load (electrophoresis of sample only) ....................... $9/sample $10/sample Short Run DNA Sequencing (reaction, purification, electrophoresis) .............................. $14.50/sample $16/sample DNA Sequencing Reaction Done (purification, electrophoresis) ......................... $10/sample $11/sample DNA Sequencing Ready to Load (electrophoresis of sample only) ....................... $7/sample $8/sample Template Preparation/Processing Template cleanup with gel and spec analysis ..................................................... $27/sample $30/sample University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 6 Plasmid Miniprep (1-5 samples) ....................................................................... $5.40/sample $6/sample Plasmid Miniprep (5 samples or more) ............................................................. $3.60/sample $4/sample Phage Miniprep ................................................................................................... $27/sample $30/sample Plasmid or phage Midiprep (1-2 samples) ........................................................... $30/sample $33/sample Plasmid or phage Midiprep (3 or more samples) ........................................... $20.50/sample $23/sample Plasmid or phage Maxiprep (1-2 samples) .......................................................... $40/sample $45/sample Plasmid or phage Maxiprep (3 or more samples) ............................................... $30/sample $34/sample Plasmid or phage Mega prep (1-2 samples) ....................................................... $72/sample $80/sample Plasmid or phage Mega prep (3 or more samples) ............................................. $57/sample $64/sample PCR amplification of template (1-2 samples) ...................................................... $11/sample $12/sample PCR amplification of template (3 or more samples) ......................................... $6.50/sample $7.50/sample PCR product purification (column) ...................................................................... $13/sample $15/sample PCR product purification (gel) .....................................................................................$22.50 $25.00 Transformation ............................................................................................................ $22.50 $25.00 Site-directed Mutation, Mutagenic Oligos not included .............................................$200.00 $225.00 Sub-cloning, DNA miniprep and Sequencing not included ........................................ $117.00 $130.00 GeneScanning / Fragment Sizing GeneScan with lane tracking and size calling (up to 48 lanes) ................................ $135/gel $150/gel GeneScan without tracking/sizing (up to 48 lanes) .................................................... $90/gel $100/gel Oligonucleotide Synthesis and Purification Note: There is no Setup Charge for Oligo Synthesis Oligonucleotide synthesis 40 nmole (0.3-0.5 OD/base) ......................................... $.85/base $.90/base Oligonucleotide synthesis 0.2 µmole (1-2 OD/base) ............................................ $1.25/base $1.40/base Oligonucleotide synthesis 1.0 µmole (5-8 OD/base) ............................................ $3.15/base $3.50/base OPC purification (5-10 OD units per column) ...................................................... $11/column $12/column Oligo w/ 5’ aminolink ............................................................................... One additional base Oligo w/ 3’ aminolink ................................................................................................. Add $20 Oligo w/ 5’ biotin ....................................................................................................... Add $15 Oligo w/ inosines ...................................................................................... Add $2 per inosine Oligo w/ 5’ fluorescent tag (FAM, HEX, TET) ........................................................... Add $20 Oligo w/ phosphorothioate bonds .......................................................... No additional charge Oligo w/ phosphorylation (3’ or 5’) .......................................................... One additional base Analysis/Assembly Training is available at no charge. selection and synthesis, sequencing, sequence evaluation, and assembly. Charges for subcloning should be discussed with facility staff. Assembly of the overall sequence, ORF analysis, and homology searches can be performed, at no charge, if requested by the user. GeneScan Identification of VNTR alleles and other sizing application can be performed on the 377 using GeneScan gels and software. Reduced charges apply if the user performs lane tracking and size calling. Charges are per gel and each gel can be loaded with 48 multiplexed samples. Sequence Assembly and analysis Assembly of sequences into a contig, identification of ORFs, analysis of properties of the predicted protein, and other analyses can be performed at the facility using MacVector and other software. These services are included at no charge for large sequencing projects, and they are available at no charge to Cancer Center and Department of Genetics members. Database Searches BLAST, BLOCKS, BLITZ, PROSITE and FASTA searches are performed using WWW or e-mail servers at no charge. Please contact the facility to arrange for database searches or for training. Genomic sequences can also be processed to identify interespersed repeats, low complexity regions, GRAIL exons, transcription factor binding sites, frequency of mono, di, and tri nucleotides, and other features. Genomic sequences can also be submitted to the GAIA University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 7 genome annotation system at the Bioinformatics Recommended Procedures for Template Preparation Evaluating your DNA template PLEASE NOTE: Criteria for cycle sequencing are more stringent than manual sequencing and many other molecular biological procedures. The success of sequencing reactions depends critically on having high purity template at the correct amount. The most important contaminants are bacterial proteins and cell wall carbohydrates, and organic solvents (ethanol, isopropanol, etc.). Please follow these recommendations for checking concentration and purity of your template DNA: • Scan DNA sample in a spectrophotometer between 220 and 320 nm. If you do not have access to a scanning spec, measure absorbance at 220, 260 and 280 nm. In Figure 1, the solid line shows the absorption profile of pure DNA (RNA profile is very similar), and the dashed lines indicate how this profile is modified by the presence of contaminants. Nucleic acids have their peak absorbance at 260, with a 260/ 280 ratio of 1.8 for DNA, 2.0 for RNA. Estimate DNA concentration from OD260: conc (ug/ml) = OD260 X dilution factor X 50. Remember that OD260 can have contributions from DNA as well as contaminants. • Run a sample of DNA on an agarose gel with a quantitative standard (e.g. lambda HindIII digest, where the amount of DNA in each band is known). By comparing intensities of template band with comparable size bands in standard lane, estimate concentration of DNA (see Figure 2). Compare this estimate to the OD260 estimate of DNA concentration. If OD260 estimate is significantly higher than gel estimate, it indicates contamination with RNA or RNA fragments. An RNase treatment, followed by phenol/chloroform extraction and precipitation would be recommended. • If the scan profile or low OD260/280 ratios indicate protein/carbohydrate contamination, perform the phenol extraction and precipitation procedure described for optional cleanup below. If organic contamination is suspected (OD220>>OD260), you can usually remove them by precipitation and redissolving. Alternatively, small contaminants can be washed away by centrifugation through a microcon or centricon ultrafiltration concentrator. Plasmids Grow 1 colony in 5-25ml LB + antibiotic overnight (~16hrs) at 37oC with shaking. Cell density should not exceed 1.5 A600 units. Debris from high-density culture inhibits the reaction. A. Purification by QIAGEN Columns Use 1.5 ml of culture lysate per mini column, 5-10 ml per midi column, or 50-100 ml per maxi column. Follow QIAGEN procedure, but add fresh RNase to 10U/ml (instead of, or in addition to, the RNase in the P1 buffer). Mix suspension very gently during and after cell lysis. Centrifuge lysate twice after lysis (supernatant should be clear). Treat with an additional 10U/ml RNase for 30min at 37oC before loading onto the column. Following isopropanol precipitation, redissolve DNA in water and precipitate with ethanol: add 1/10 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 8 volume 3M sodium acetate + 3 volumes absolute ethanol, chill on ice 30 minutes. Spin at 10,000 RPM for 20 min, 70% ethanol wash, dry, redissolve in water (salts inhibit sequencing reaction). B. Purification by Promega Wizard™ Mini Preps Use 3ml of culture per mini prep and follow Promega procedure. Spin lysate twice. Be sure the supernatant is clear before adding resin. It is often necessary to concentrate eluted samples by ethanol precipitation or drying. Precipitation also improves quality of template FIGURE 1 Spectroscopic analysis of DNA sample and allows resuspension in water. C. Optional Purification 1. RNase treatment, followed by phenol/chloroform extraction and precipitation improves template quality, but care must be taken to remove all organics. Most samples treated in this manner can be sequenced successfully. Incubate with 10 U/ml fresh RNase for 30 min at 37oC, extract with phenol (until interphase is clear), phenol/chloroform, then chloroform. Precipitate with ethanol, wash pellet with 70% ethanol, dry, and redissolve in water. 2. Template quality may also be improved using a 100K centrifugal concentrator (minicon, centricon – dilute 5-10 µg of DNA to 50-100 µl with water and centrifuge through concentrator at 3000g for 10 min, add water and repeat 3-5X) or by dialysis for 2hr using an 100K MW cut off Millipore Minitan-S membrane (float the dry membrane shiny side down on sterile water and apply the 20 µl DNA sample). 3. PEG precipitation is also helpful in improving quality of template. Add 0.25 vol 4M NaCl and 1.25 vol 13% PEG 8000, incubate on ice 20 min, pellet DNA, redissolve in water. D. Alternative methods Templates purified by phenol-chloroform extraction or CsCl gradients can be used, as long as sufficient further precipitations/washes are done to remove organics/CsCl. PCR Products If a PCR product produces a single band, purify with QIAGEN or Wizard™ PCR purification columns. These columns use salts and ethanol, so extensive washing of pellet and complete drying of ethanol is necessary. Several washes with water through a microcon 100 concentrator can also be used for purification or clean-up. If there are multiple bands, run products on low melting point agarose gel, excise band, purify DNA using QIAGEN QIAEX II or Promega Wizard™ PCR preps following recommended procedure. Alternatively, run products on a gel, cut and electroelute desired band. Quantitate template on gel (OD260 can be misleading if salts/alcohols are left in sample). Submit 10ng of DNA / 100 bp / reaction in = 5 µl water. Bacteriophage Lambda Lambda preps isolated by two consecutive bandings in CsCl, followed by RNase treatment and phenol-chloroform extraction, yield good DNA for sequencing. Purification by QIAGEN Lambda prep columns also gives adequate University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 9 results. Below is a procedure for preparing lambda DNA by extractions and precipitations. 1. Place 200 µl of a healthy overnight culture of the appropriate host strain into a clear plastic, sterile 10 ml snap-cap tube. The choice of bacterial strain is important. Choose a host in which lambda replicates vigorously, e.g. use strain Y1090 for a lambda gt11 clone. If appropriate, include 0.2% maltose in culture medium to induce the lamB gene that codes for lamba receptors. 2. Stab a well-separated plaque with a sterile toothpick and rinse the captured lambda phage particles into the cells aliquoted above. Alternatively, use 3X10**6 pfu of phage from a stock. Allow the phage to adsorb to the cells for 15 minutes at room temperature. 3. Add 2 ml Tryptone Broth containing 10 mM MgSO4 and shake the culture at 37oC until the culture is lysed. Lysis is evident when the culture becomes clear and cellular debris is prevalent. This will take anywhere from 4 to 8 hours depending on both the host and lambda strain used. Add 10-100 µl chloroform and continue shaking to complete lysis if necessary. Pellet cellular debris at 10,000 rpm for 5 min. 4. Pour the supernatant into a new 2 ml tube. Add 10 µl 2000U/ml RNase A and 5 µl 10 mg/ml DNase I. Incubate the sample at 37oC for 20 minutes. 5. Add 0.12 g NaCl and invert the tube several times until the salt is dissolved, then add 0.2 g polyethylene glycol (8000 average MW) and invert the tube repeatedly until the PEG is dissolved. Let the sample sit at 4oC for 1 hr to overnight. 6. Spin in a microcentrifuge for 20 minutes. Resuspend pellet in 0.5 ml SM (50 mM Tris, pH 8.0, 100 mM NaCl, 10 mM MgS04) and transfer to a 1.5 ml tube. 7. Extract the sample once with chloroform. To the aqueous layer, transferred to a new tube, add: 40 µl 250 mM EDTA pH 8.0, 5 µl 20% sodium dodecyl sulfate, 2.5 µl of 10 mg/ml Proteinase K, and incubate the sample at 60oC for 30 minutes. 8. Extract twice with phenol/chloroform, then twice with chloroform. Add 40 µl 3 M sodium acetate, pH 5.5 and FIGURE 2: Gel analysis of DNA sample In this example, 1 ul of lambda HindIII digest containing 500 ng DNA was loaded in the outer lanes, and 10 ul of sample was loaded in lanes 2 and 3. Since the lambda genome is 50 kb in size, the 4.4 kb band contains 500 X 4.4/50 = 44 ng of DNA. The sample in lane 2 is a little brighter, probably containing 80 ng of DNA. Therefore, DNA concentration in the sample is 8 ng/ul. 1ml 95% ethanol. Spin at room temperature for 10 minutes. 9. Rinse the pellet with 750 µl cold 70% ethanol. Resuspend the DNA pellet in 400 µl distilled water. 10. Add 200 µl 40% PEG8OOO, 30 mM MgSO4. Mix thoroughly, let stand at room temp for 20 min. 11. Extract twice with chloroform. Add 40 µl 3 M sodium acetate and 1 ml 95% ethanol. Mix thoroughly. Spin at room temp for 10 min. 12. Rinse the pellet with 750 µl cold 70% ethanol. Air dry pellet and resuspend in 50 µl distilled water. Yield is 12 µg. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 10 BAC DNA purification Recently QIAGEN and Princeton Separations have introduced their BAC DNA kits. We have mixed results with both. We describe here the NH4Ac/QIAGEN midi prep method. BAC DNA prepared by CsCl gradient centrifugation, followed by Rnase treatment, extraction and precipitation work well as templates. Below is a modified alkaline lysis method that yields good quality DNA. 1. Grow 200 ml of culture. OD 600 should be between 1 and 1.5. 2. Spin down cells and resuspend in 15 ml of P1 adding fresh RNase (150 units). 3. Add 15 ml of P2, mix by gentle inversion and incubate for 5 minutes at room temperature or until lysis looks complete. 4. Add 15 ml of P3 and keep on ice for 15 minutes. Mix every 3 minutes by gentle inversion. 5. Spin down cell debris 13,000 rpm for 30 minutes at 4 C. Spin a second time for 15 minutes to get rid of any debris still in suspension. 6. Transfer supernatant to a fresh tube and precipitate with isopropanol. Wash pellet with 70% ethanol. 7. Resuspend pellet in 1 ml H2O and add 0.25 ml 10 M NH4Ac to a final concentration of 2 M NH4Ac. Pellet precipitate by centrifugation at 13,000 rpm 30 min, keep supernatant. 8. Transfer supernatant to a new tube and precipitate DNA with 2 volumes of ethanol. Pellet at 10,000 rpm for 15 minutes. 9. Redissolve pellet in 2 ml of H2O and add 10 ml of QBT. 10. Equilibrate QIAGEN tip 100 with 4 ml of QBT. Apply sample to column. 11. Wash column with 10 ml of QC three times. Elute DNA with 4 ml of QF. 12. Divide eluted DNA into 6 microfuge tubes, 0.67 ml each. Add 0.670 ml of isopropanol and chill on ice for 15 min. 13. Spin DNA down at 10,000 rpm for 15 min and wash pellet with 70% ethanol as before. 14. Dissolve in water after air drying. Yield is 5-8 µg. Dye Terminator Cycle Sequencing Generally follow protocol given by ABI for Taq FS and Big Dye™ terminator chemistry. Consult ABI protocols or facility staff for other chemistries. Typically, template is in 6 µl volume, primer in 3 µl, and reaction mix with Taq polymerase in 4 µl. Add DMSO to 3%, and double Taq concentration, for GC rich or difficult templates. Total reaction volume is 13 µl. Table 3 shows template, primer and reaction condition requirements for different templates when using Big Dye™ terminators and Taq FS: Reaction products are purifed with AGTC 96 Well Gel Filtration Block (Edge Biosystems). Purification can also be done with Centrisep Spin columns (Princeton Separations) or a Sephadex G-50 Quick Spin column from BoehringerMannheim, or Centrisep column washed and repacked with G-50. AGTC 96 Well Gel Filtration Block Variable speed centrifuge equipped with a rotor and a microplate carrier is required. Follow manufacturer’s instructions. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 11 CentriSep Swell dry gel with 0.75 ml of water for 1hour. Spin in fixed angle microfuge rotor at 750g (~3,000 rpm) for 2 min. Put tubes in with mark oriented up, so that you can perform second spin in same orientation. Load sample in center of gel, repeat spin with tube in same orientation as the first spin. BMB Spin Columns If using an angle rotor, orient the columns the same way for both spins. After the first spin, the gel material within the column will pull away from the column walls. Be sure to load your sample in the center of the gel material. Dry down sample in SpeedVac. Resuspend in 2.0 µl of loading buffer, denature for 2 min at 95 °C (DO NOT OVERHEAT!), keep on ice, load. Loading buffer: 5 parts deionized formamide + 1 part 25 mM EDTA, 6% blue dextran, pH 8.0 TABLE 3: Amounts of template and primer to be submitted (Please add primer to your template in the suggested volumes) Template type Size (kb) Template/rxn Suggested concXvol Primer amount Primer conc. PCR product 0.1-1 10 ng per 100 bp 1.6-16 ng/µl X 6µl 3.2 pmoles 1.1 µM X 3µl Plasmid 2-15 0.5 µg 80 ng/µl X 6µl 3.2 pmoles 1.1 µM X 3µl Phage/cosmid/P1 20-100 1.0 µg 110 ng/µl X 9µl 12 pmoles 4 µM X 3µl BAC 200 2.0 µg 220 ng/µl X 9µl 12 pmoles 4 µM X 3µl Network Connections A. You can communicate with the sequencing facility, receive information, and order oligos by e-mail: Tapan Ganguly: ....................dnaseq@mail.med.upenn.edu Debi McKinney .................. dlmckinn@mail.med.upenn.edu Peiqui Chen .........................pchen2@mail.med.upenn.edu Elias Papaioannou ............... eliasp2@mail.med.upenn.edu Lanping Zhang .................... lpzhang@mail.med.upenn.edu David Zhu .................................zhu2@mail.med.upenn.edu B. Access for all Macintosh users: Macintosh computers running System 7.6 or later can use AppleShare to connect to our server over the Internet, even if they are not connected directly to PennNet. (You may need to update your AppleShare software to do this, and we urge you to contact your Local Support Provider for more information. Updates can be downloaded from http://asu.info.apple.com/swupdates.nsf/artnum/ n11419.) You will need to contact the facility and choose a user/ folder name and a password for your folder. We will put your results in your folder to which only you have access. Because of the large number of users and various users with the same investigator, you must indicate on each Sequencing Request Form that you want results put into a folder, and give the folder name. Results will be kept in your folder for up to one month. All results are also archived by the facility if you need them. Note that the purpose of the server is to enable you to copy files and disconnect when done. Please do not doubleclick the files on the server! 1. Select “Chooser” from the Apple menu. 2. Click on the AppleShare icon in the upper-left corner, then click on the “Server IP Address…” button near the lower-right corner. 3. In the input box which appears, type in “dnaseq.med.upenn.edu” and click “Connect”. 4. When prompted, type in your lab folder name and University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 12 password and click “OK” or “Connect”. 5. In the next box that appears, “Sequencing Results” should already be selected. Do not click the checkbox on that top line, as this may slow down the server’s performance and/or cause your computer to crash. Just click “OK”. 6. Read the current update message that appears and click “OK”. 7. Close the Chooser window, and you should find that the “Sequencing Results” server volume is now on your desktop. On future occasions, you can skip the above steps by selecting “Sequencing Results” from the “Recent Servers” sub-menu under the Apple menu. Your results will be located in your lab folder on the “Sequencing Results” server volume. Please note that results will be available after 11:00 a.m. on most days. Copy the files to your computer, then disconnect from the server by dragging the “Sequencing Results” icon to the Trash. Near the top of the folder list, you will also find installers for Sequence Editor and EditView software for examining sequence data, as well as Sequencher and ClustalW software for multiple alignment of sequences. Be sure to copy these items to your own hard drive rather than installing the software directly from the server. C. Other Internet Access: Anyone with access to the Internet can copy their results by FTP, using Fetch for Macintosh, WS-FTP for Windows or other file transfer software. To do this, set up a user name and password, as in section B above. Connect to the FTP site “dnaseq.med.upenn.edu” using your lab folder name and password. If your software has a space for specifying the directory, leave it blank. Once connected, find your lab folder in the directory listing and open it. Select the files you want and transfer them to your computer. Please disconnect from the server when finished copying. Guide for Oligonucleotide Synthesis Introduction Oligonucleotides are synthesized on the ABI 394 Synthesizer, using solid-phase synthesis and phosphoramidite nucleoside chemistry. Synthesis proceeds from the 3’ to the 5’ end. The 3’ nucleoside is already coupled to the solid support (polystyrene or glass), and activated nucleotides are sequentially added on to this nucleoside. Bases and phosphate groups are kept protected until the end of synthesis. At each step, the sugar protective dimethyl trityl (DMT) group on the deoxyribose is removed to allow reaction with the next amidite. Since these reactions are only ~99.5% efficient, a capping reagent is used to cap unreacted sugars. This cap terminates synthesis on the small proportion of oligos in each step that did not receive the correct next base. Because of this cap, syntheses will usually produce some terminated chain oligos, but no significant amounts of oligos with incorrect (e.g., missing internal base) sequence. Thus, the mixture of oligos will all start with the same 3’ end, but some proportion of the chains will be terminated early. TABLE 4: Yields of oligonucleotides >(20-mer) at different scales OD260 Yield (mg) Yield (nmoles) Conc. (suspended in 1 ml) 40 nmole scale 5-8 0.25-0.4 40-65 40-65 µM University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 13 0.2 µmole scale 20-40 1.0-2.0 160-320 160-320 µM Yield & Efficiency Printouts provided with oligos give two numbers that allow you to evaluate the success of synthesis: the Average StepWise Yield (ASWY), which give the average efficiency of incorporation of nucleotides at each step, and the Overall Yield, which represents the percentage of total chains that are full-length. Typical values of ASWY and overall yield in a successful synthesis of a 20-mer are 98.5% and 70%, respectively. Lower overall yields of fulllength product are obtained with longer sequences. Average yields per base for 40 nmole and 0.2 µmole scales are 0.3-0.4 OD260 and 1.0-2.0 OD260 units, respectively. The values for an average 20-mer (MW ~ 6,000) suspended in 1 ml are listed in Table 4. Purification A typical synthesis generates a mix of full-length oligos (which have the DMT group at their 5’ ends), and shorter oligos (which are capped). Oligos synthesized in this manner can be used for PCR and sequencing without OPC purification because they have a well-defined 3’ ends (the site of elongation by polymerases). Most PCR and sequencing primers therefore do not require OPC Purification. If the oligo is to be used as a linker or adaptor and the integrity of the 5’ end is also essential, then we recommend that you choose “OPC Purify”. An affinity column (OPC) that recognizes the DMT group is used to purify full-length oligos from the mix. Each OPC column can be loaded with 5-10 OD of crude oligonucleotides (which corresponds to the yield of a typical 40 nmole synthesis), and will yield 2-5 OD of purified product. The DMT group is removed during purification. For purification of larger scale syntheses, please indicate if you want purification of 10 OD only (one column), or all of the produced oligo (multiple columns). Please note that efficiency of full-length oligo production decreases with length of oligo, and 0.2 µmole scale synthesis would be recommended for long (>40) oligos that need to be OPC purified. Modifications Oligos phosphorylated at either 3’ or 5’ ends can be synthesized using the ABI phosphalink reagent. We can also add a 3’ or 5’ amino group, 5’ biotin, or a 5’ fluorescent dye (FAM, HEX or TET). Furthermore, phosphorothioate oligos can be synthesized for antisense or other applications. These modifications should be indicated on the request sheet or discussed with the staff. Please note that 5’ phosphorylated oligos do not have a DMT group and cannot be OPC purified. Evaluating and Editing Sequence Results Please do not start to work with the sequence text file before you examine the chromatogram. Sequence calls by the analysis software can have several types of problems, and we recommend that all users evaluate and edit the automated sequence calls as necessary by examining the actual colored peaks in the chromatograms. The areas that you should pay particular attention to are: 1. Errors in base calls at the start of the sequence due to noise. You may decide to trim away the initial bases University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 14 (10-20 bases). 2. Occasional bases missed in the first 50 bases due to compression. 3. Possible miscalled bases in areas where signal may be low. 4. Incorrect base calls beyond 600 bases where resolution is deteriorating. 5. Determining where usable sequence stops, and trimming the remaining base calls. Three programs are available for viewing and editing chromatograms: EditView, SeqEd, and Sequencher. You can copy EditView and SeqEd from the Server, or download them by clicking here. Sequencher is a commercial software by GeneCodes, but the facility maintains network sharable copies. EditView allow opening, editing, exporting, and printing of one sequence file at a time. It is self explanatory, and easy to use. FIGURE 3: A schematic diagram of solid phase oligodeoxynulceotide SeqEd allows opening multiple chromatograms if comparison between them is desired. SeqEd is not a good assembly program and should not be used for sequence assembly. Instructions for using SeqEd are given below. Sequencher is a sophisticated, commercial software for sequence editing and assembly. It would require significant effort to learn. If you are interested in using Sequencher, ask the facility staff for a copy of the software. In addition, copy the KeyServer extension from the Sequencher folder on the server into your system folder, and follow the instructions for how to run Sequencher using the network KeyServer. Sequencher has an on-line help function that explains the major features of the program. Short Instructions for SeqEd SeqEd allows you to examine chromatograms and the associated sequence text file, correct errors, and extract the reliable portion of the sequence from your results. We strongly recommend that users of DNA sequencing services review the chromatograms and edit the base calls made by the ABI analysis software as needed. The software sometimes makes obvious errors, particularly at the beginning and near the end of a run. SeqEd also allows you to open two or more chromatograms at the same time for purposes of comparison. To use SeqEd, follow these steps: 1. Double click on the program file SeqEd v1.0 to start it. 2. From the bar menu, select Sequences and then select Import (or use the shortcut Apple-M). A window will open up to allow you to choose the appropriate folder and chromatogram file. Do not try to open a “.seq” sequence text file. Select the chromatogram file and open it. At this time only the sequence text will show. Note that only the text part of the file is visible at this time. 3. Repeat step 2 above to select additional chromatogram files if desired. More than three chromatograms per screen tends to be too crowded. 4. Select the SeqEd window that contain all the text information of the files you imported, then select the individual file names you want to visualize. You can use shift-click to select multiple files. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 15 5. From the Sequences menu, select Display Chromatogram (or use the shortcut Apple-D). All selected chromatograms will appear on the screen in compressed mode. If necessary, select specific files and then select Reverse Complement Sequence from the Sequences menu. 6. Select each chromatogram one at a time, click on the “C” button in the control panel on the bottom left, then click the pointer in an area near the beginning of the chromatogram. The chromatogram will expand to make peaks and the sequence visible. 7. Scroll along the chromatogram, looking for areas where the base calls by the analysis software might be in question. Look for missing bases, extra bases, peaks that were called as N’s when they should not have been (signal-to-noise > 2), or peaks that were called as a base but should be N (signal-to-noise <1.5). Click on the base or position you want to edit, and type in the new base. 8. Decide where you think the reliable sequence ends. Use both signal-to-noise and peak broadening to determine the last reliable base. Click on it to determine what base number it is. 9. Click on the top window containing the text sequence, select the specific sequence you have just edited (if working with multiple sequences), find the location of the last reliable base, and delete the remainder of the sequence. 10. From the Sequence menu, select Export (or use the shortcut Apple-E). A window will appear to allow you to choose the location of the file, its name, and what type it is. Select Text, and change the name of the file (you are not allowed to overwrite an original sequencing result file). The edited text sequence will be stored in the selected file and location. 11. This text file can be opened in word processors or other sequence analysis software for further manipulation and analysis. 12. When finished, quit SeqEd and answer No to all the questions it asks before closing. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 16 CELL CENTER SERVICES FACILITY David Gasser, Ph.D., Director 215/898-5175 gasserd@mail.med.upenn.edu Tapan Ganguly, Ph.D., Technical Director 215/573-7238 gangulyt@mail.med.upenn.edu Sabine Baxter, Research Specialist 215/898-2795 baxters@mail.med.upenn.edu Diane Miller, Research Technician 215/898-2795 dianemil@mail.med.upenn.edu Location: D2 Richards Building Mailing Address: D2 Richards Building 3700 Hamilton Walk Philadelphia, PA 19104-6085 Telephone: 215/898-2795 Fax: 215/573-9327 Internet: http://www.med.upenn.edu/ccsf/ Consultation • Training • Cell bank and Seed Cultures • Liquid Nitrogen Freezer Storage • Lymphocyte & DNA Banking • EBV Transformations • Large Scale Cell Culture • Mycoplasma, Endotoxin and MHV Testing • Specialty Media • Autoclaving • Hybridoma Production and Screening • Antibody Production (supe or ascites), Purification & Conjugation • Cancer Center Shared Resources and Facilities U N I V E R S I T Y O F P E N N S Y LVA N I A D E PA R T M E N T O F G E N E T I C S Cell Center Services _ Please fill out a “Request for Service” form when using Cell Center Services _ Please acknowledge services performed by the facility in your publications 1. Consultation The Technical Director will discuss projects with investigators to explore and evaluate different experimental designs and approaches utilizing cell culture and immunological techniques. Establishment and use of cell lines, strategies for production of monoclonal antibodies, and other options can be explored with investigators. 2. Personnel Training The Cell Center will train laboratory personnel in basic sterile and tissue culture techniques, hybridoma and University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 17 monoclonal antibody production procedures, and immunological assays. An individual training program in basic cell culture skills, which runs for a total of 12 hrs over a two week period, is offered for a charge of $300. This program will expose inexperienced personnel to basic techniques of cell culture, including making media, thawing cells, cell count and determination of viability, culture and subculture of attached cells and cells in suspension, and freezing cells. We also have available two 35 min videos and some worksheets on cell culture techniques. These can be signed out by users for 2-3 days at no charge. They are useful for understanding basic sterile culture techniques, or as a refresher for more experienced people. Please contact us to arrange for training in more specialized areas. 3. Seed Cultures (established lines from our cell bank) 1. T-25 ......................... $30.00 2. T-75 ......................... $35.00 3. T-150 ....................... $40.00 We can also order ATCC cell lines not available in our cell bank (usual cost ~$120). 4. Preparation of Special Media The Cell Center prepares and sterilizes custom formulations, as long as a commercial powder base is available as a reasonable starting point. Pricing for special order media is as follows: $20.00/liter for orders less than 10 L $15.00/liter for orders 10 L or more $13.00/liter for orders 10 L or more, and powder provided by user 5. Drosophila Culture Medium The Cell Center prepares media for Drosophila culture in vials and bottles. Charge for a tray of 100 vials is $18.00; a tray of 25 bottles costs $20.00. 6. Endotoxin Testing We will test water or samples of media you have prepared to determine endotoxin levels, using the LAL (Lumilus Amebocyte Lysate) gel clot assay. At least 200 µl of sample should be submitted in an endotoxin free container. Generally, radiation sterilized tissue culture plasticware is endotoxin free, but autoclaved glassware is not. Three dilutions per sample are tested routinely, along with a set of blanks and positive controls. Please indicate desired dilutions on the request sheet. Prices for tests are: $15.00/sample for 1-2 samples, $10.00/sample for 3 or more samples. There will be additional charges for testing blood or serum samples because of time consuming boiling and clarification steps required. 7. Mycoplasma Testing: _ For the protection of personnel performing tests, please clearly label supernatants from human cell lines and virally infected or transformed cell lines. _ Mycoplasma testing of cell lines that are to be introduced into animals is strongly recommended by ULAR. MHV testing is recommended for lines passaged through, or to be introduced into mice. Both tests are available at the Cell Center Service Facility. 7A. Positive/negative verification using fluorescent DNA staining: $10.00 per sample ($20.00 per sample for non-University of Penn. investigators) Requirements University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 18 are as follows: a) All lines must be antibiotic-free for at least two passages. Presence of antibiotics can give false negative results. b) Submit 1-2 ml of sterile supernatant (no cells) in a 15 ml screw cap tube. Presence of cells or cell debris can produce false positive results. Supernatant has to be in contact with the cells for at least 48 hours. c) All samples must be submitted by Noon on Thursday, along with a request card. d) Test results will be called in on the following Tuesday or Wednesday. 7B. PCR assay for mycoplasma: The Cell Center will perform a PCR test based on a set of five primer pairs that produce unique fragments from the five most common strains of mycoplasma (M. orale, M. hyorhinis, M. fementans, A. laidlawi, M. arginini). Products are analyzed by agarose gel electrophoresis. Sample requirements are 0.2-0.5 ml of sterile culture supernatant from cells grown without antibiotic. Cost of the test is $25/sample for 1-2 samples, $20/sample for 3 or more samples. 8. Mouse hepatitis virus testing: Cultured cells (e.g. hybridomas) that are passaged through or intended to be introduced into mice will be tested for mouse hepatitis virus by RNA extraction from culture supernatant and cell pellet, followed by RT-PCR. Please provide a viable culture in a T25 flask. ULAR also recommends that such lines be tested for mycoplasma (see above). Cost of the RT-PCR test on both supernatant and cell pellet is $100. 9. Lymphocyte isolation, EBV Transformation, DNA extraction Lymphocytes from patient blood samples are isolated by centrifugation over Ficoll. These lymphocytes are frozen and/or transformed by EBV and cultured until 3 vials can be frozen. Charge for lymphycte isolation and freezing is $50/sample. Charge for EBV transformation is $180. DNA can also be extracted from blood, lymphocytes, or cultured EBV transformed cells for a charge of $30. Contact facility for cost of larger scale cultures, and reduced charges for large number of samples. 10. Roller Bottle and Spinner Flask Facilities Available free of charge if your personnel do the work, or Cell Center personnel will do the work for a negotiated fee. 11. Liquid Nitrogen Storage The Cell Center rents space in liquid nitrogen freezers at the rate of $1.00 per slot per quarter. 12. Autoclaving The Cell Center will autoclave clean glassware and properly packages biohazard waste at a charge of $5.00 per cycle. 13. Hybridoma Production and Screening The Cell Center will assist in monoclonal antibody production, including consultation and development of approaches appropriate to each research project, antigen preparation, immunization, fusion, screening, and cloning. We use a high efficiency fusion protocol that results in at least 1000 hybridoma containing wells University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 19 per fusion. Typically, the Cell Center will perform immunizations, fusion, collection of supernatants, and subcloning of up to 20 positive lines for $2,000, while a fusion involving subcloning of up to 40 positive lines will cost $3,000. An additional $1,200 is charged for an ELISA screen. Please see the “Fusion Request form” and contact the Cell Center staff for more information. 14. Production and Purification of Antibodies The Center will produce and purify antibodies from culture supernatant or ascites. Hybridoma cells can be grown either in roller bottles or in hollow fiber cartridges. Charge for a liter of culture supernatant grown in roller bottles is $300; concentration by precipitation and purification of an IgG antibody by protein-G chromatography through FPLC cost $200.. We also do affinity purification of antibody against a specific antigen; the cost of making the affinity column is $100 (1 ml) or $150 (5 ml), the cost per run is $100. If protein-free supernatant is required, a slow transition to protein-free medium will be undertaken for a charge of $200. Charges for hollow fiber culture are as follows: the set up charge $450, charge per week $40 (0.1 - 0.5 g of antibody, depending on the hybridoma line). Charges for ascites production are $100 per mouse. Please note that some hybridoma lines will not grow as ascites. Please consult with facility staff if the line has not previously been grown as ascites. Typical yield of successful ascites procedures is 3-6 ml per mouse. ULAR strongly recommends that cells to be used for ascites be tested for mycoplasma and mouse hepatitis virus (see services #7 and 8 above). 15. Antibody isotyping, fragmentation, conjugation Murine monoclonal antibodies are isotyped with a commercial kit utilizing subclass specific antibody reagents. Charge for isotyping is $20 per line. Fab and F(ab)2’ fragment can also be produced by the facility, as well as FITC, biotin, or enzyme conjugation of purified antibody. 16. Anti-myc antibody The facility provides protein G purified, ELISA tested myc-1 monoclonal antibody from the 9E10 hybridoma for use by University investigators at a cost of $30/mg. 17. ELISA reader (Titertek’s Multiscan MC, in room D2 Richards). Free of charge, and available to all members of the University community, 9:00 a.m. - 4:30 p.m. 18. Cell Line Searches The Cell Center has access to ATCC cell line and hybridoma databases. The Cell Center will perform searches for cells in these databases at your request free of charge. 19. Other Shared Resources The Cell Center is a shared resource and hybridoma core facility for the Cancer Center. As much as practically feasible, Cell Center equipment is available for Cancer Center members’ use, particularly in pilot/ start up projects. This equipment includes: • Sterile culture hoods, incubators, and accessory equipment • Roller bottle and spinner flask facilities • Centrifuges University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 20 • Protein and DNA electrophoresis systems, electrophoretic and positive pressure blotters • UV cross-linker, transilluminator • Speedvac concentrator and gel dryer • Low pressure liquid chromatography system • Zeiss fluorescence microscope and camera • Leica inverted microscope and camera • Sonic disruptor • PEC DNA Cycler 480 • Zeiss 505E cryomicrotome Summary Charge for Services Charge for Cancer Center Service Description General Charge & Genetics Antibody isotyping (per line) $30.00 $28.00 Antibody purification (protein G) $200.00 $180.00 Antibody purification Affinity column (1 ml) $100.00 $90.00 Affinity column (5 ml) $150.00 $135.00 Cost per run $100.00 $90.00 Antibody conjugation $100.00 $90.00 Ascites (per mouse) $100.00 $90.00 Autoclave (cycles) $5.00 $4.50 Cell culture Large scale (1 L) $300.00 $270.00 Cell culture Large scale (hollow fiber) Set up charge $450.00 $405.00 Charge per week (0.1-0.5gm of antibody) $140.00 $126.00 Cell Culture T150 $55.00 $49.50 Cell Culture T75 $45.00 $40.50 Cell Culture T25 $40.00 $36.00 DNA isolation (10-20 ml blood or 100 ml culture) $30.00 $27.00 Drosophila medium (100 vials) $25.00 $25.00 Drosophila medium (25 bottles) $25.00 $25.00 EBV stock $6.50$6.0 0 Endotoxin test (1-2 tests per test) $15.00 $13.50 Endotoxin test (3 or more tests per test) $10.00 $9.00 Fusion U Penn 20 clones $1900.00 $1710.00 Fusion U Penn 40 clones $2900.00 $1610.00 Fusion non-U Penn 20 clones $2600.00 Fusion non-U Penn 40 clones $4000.00 Fusion Screen ELISA $1200.00 $1080.00 Liquid N2 storage (per vial/quarter) $1.00 $0.90 Lymphocyte EBV Transformation $180.00 $162.00 Lymphocyte isolation $50.00 $45.00 MHV test RT-PCR $100.00 $90.00 Myc-1 antibody from 9E10 (1 mg) $30.00 $27.00 Mycoplasma test PCR (1-2 samples per sample) $25.00 $22.50 Mycoplasma test PCR (>3 samples per sample) $20.00 $18.00 Mycoplasma test Hoescht non-U Penn (per sample) $20.00 Mycoplasma test Hoescht U Penn (per sample) $10.00 $9.00 Personnel Training cell culture $300.00 $270.00 Phage Secreted scFv production (1L) $250.00 $225.00 Special Media per L (10L or more powder provided) $13.00 $11.70 Special Media per L (10L or more) $15.00 $13.50 Special Media per L (less than 10L) $20.00 $18.00 Transition to protein free medium (per line) $200.00 $180.00 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 21 GENETIC DIAGNOSTIC LABORATORY Haig H. Kazazian, Jr., M.D., Medical Director 215/898-3582 kazazian@mail.med.upenn.edu Arupa Ganguly, Ph.D., Director, Research and Development 215/573-3020 ganguly@mail.med.upenn.edu Catherine Stolle, Ph.D., Managing Director 215/573-3030 stolle@mail.med.upenn.edu Lynn Godmilow, M.S.W., Director, Genetic Counseling 215/573-9161 godmilow@mail.med.upenn.edu Location: 415 Anatomy Chemistry Building 339 Anatomy Chemistry Building Mailing Address: 502 Clinical Research Building 415 Curie Boulevard Philadelphia, PA 19104-6145 Telephone: 215/573-9161 Fax: 215/573-5940 Internet: http://www.med.upenn.edu/gdrs Genetic Diagnostic Laboratory About the Genetic Diagnostic Laboratory The Genetic Diagnostic Laboratory in the Department of Genetics is pleased to offer state-of-the-art DNA Diagnostic Testing and consultation for a wide variety of genetic diseases and conditions. Pre-test genetic counseling, rapid communication of results and accurate interpretation with recommendations are available. For more information about the available diagnostic tests, sample requirements, and the protocols for requesting tests, please contact Lynn Godmilow, MSW, CGC, Genetic Counselor and Director of our Genetic Diagnostic Referral Service by telephone (215/573-9161), fax (215/573-5940), or by email (godmilow@mail.med.upenn.edu) Available Molecular Tests for Genetic Disorders Adenosine Monophosphate Deaminase-1 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 22 (AMPD1) Background: Deficiency of this muscle specific enzyme is a common cause of exercise-induced myopathy characterized by weakness and cramping and probably the most common cause of metabolic myopathy in humans. Assay: Direct mutation by PCR Utility: To detect carriers and affected individuals as part of the differential diagnosis of exercise induced myopathy CPT codes: 83890, 83892, 83894, 83898, 83912 Turnaround: 1 week •-Thalassemia Background: Autosomal recessive disease found mostly in Southeast Asians caused by deletion of aglobin genes with severity dependent on the number of deleted genes and leading to lifelong dependence on transfusions with iron buildup and shortened life-span. In AfricanAmericans deletion of fewer alpha genes than in Southeast Asians can be clinically similar to ß-thalassemia trait. Assay: Direct mutation by Southern blot. Direct detection of Hb Constant Spring if indicated Utility: Confirmation of diagnosis, carrier detection, prenatal diagnosis CPT codes: 83891, 83892, 83894, 83896, 83897, 83898, 83904, 83912 Turnaround: 2-3 weeks ß-Thalassemia (Cooley’s Anemia, Mediterranean Anemia) Background: Autosomal recessive disease, common in the Mediterranean, Asian, Indian, Middle Eastern and African-American populations, caused by decreased or absent ß-globin chains; characterized by severe anemia, growth retardation, splenomegaly, bone changes with characteristic facies and iron depositions affecting cardiac and endocrine systems. Assay: Direct mutation by PCR or multiple PCR’s Utility: Confirmation of diagnosis, carrier detection, prenatal diagnosis CPT codes: 83890, 83891, 83892, 83896, 83897, 83898, 83904, 83912 Turnaround: 2 weeks CADASIL Background: CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) is characterized by history of migraine headaches, mid-adult (30s-60s) onset of cerebrovascular disease progressing to dementia, and diffuse white matter deficits on neuroimaging. CADASIL is caused by mutations in the Notch3 gene (chromosome 19).tems. Assay: Direct mutation analysis by Conformation Sensitive Gel Electrophoresis (CSGE). Analysis of PCR products for the presence of heteroduplexes followed by sequencing to detect differences from the consensus sequence of Notch3. There are three levels of testing performed. Level 1 consists of University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 23 commonly mutated exons and will detect 87% of mutations in skin biopsy proven cases. Level 2 consists of testing all other exons in which a mutation has been reported and will improve sensitivity to 93% in biopsy proven cases. Level 3 consists of testing the remaining exons encoding EGF like domains and will be performed for a limited time on a research basis at no additional cost to the patient. There is currently no information regarding the increase in sensitivity (if any) with Level 3 testing. Utility: Diagnostic confirmation, prenatal diagnosis, presymptomatic testing (presymptomatic testing is available by special arrangementtalk with the genetic counselor) CPT codes: Level 1- 83890, 83898, 83898-76, 83903, 83904, 83904-76, 83912; Level 2 - 83898, 83898-76, 83903, 83904, 83904-76 Turnaround: 2 weeks for Level 1; 2weeks for Level 2 Charcot-Marie-Tooth Disease (X-linked) Background: Sex linked dominant neurodegenerative disorder . Assay: Sequencing of the connexin 32 gene detects mutations in >85% of patients. Utility: To confirm the clinical diagnosis and to offer prenatal diagnosis CPT codes: 83890, 83898, 83894, 83904, 83912 Turnaround: 4 weeks Craniosynostosis Syndromes: Apert, Coronal Synostosis (Plagiocephaly), Crouzon, JacksonWeiss, Pfeiffer Background: Highly penetrant, autosomal dominant disorders characterized by premature fusion of skull bones leading to an abnormal head shape, ocular hypertelorism with proptosis, midface hypoplasia, and the presence or absence of characteristic hand and/or foot manifestations. Although many cases are sporadic, familial cases have been reported. Diagnosis is possible on the basis of clinical exam in many cases, however, variable expressivity can result in ambiguity or misdiagnosis and, prenatal diagnosis based on ultrasound is unreliable. These disorders are due to mutations in fibroblast growth factor receptors (FGFR1, 2, 3). Assay: Direct mutation detection by restriction enzyme digestion or sequence analysis of PCR-amplified genomic DNA. Utility: Confirm a presumed clinical diagnosis or for prenatal diagnosis in familial or sporadic cases. CPT Codes: Apert: 83890, 83892, 83894, 83898, 83912 Coronal Synostosis: 83890, 83892, 83894, 83898, 83912 Crouzon: 83890, 83894, 83898, 83904, 83912 Jackson-Weiss: 83890, 83894, 83898, 83904, 83912 Pfeiffer: 83890, 83892, 83894, 83898, 83904, 83912 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 24 Turnaround: 1-2 weeks Duchenne/Becker Muscular Dystrophy (DMD/ BMD) Background: Duchenne Muscular Dystrophy is an X-linked progressive neuromuscular disorder beginning in childhood with muscle weakness followed by wheel chair dependence and leading to a shortened life span. The defect has been shown to reside in the gene coding for a protein called dystrophin. The milder Becker form, also X-linked, is less common, has later onset with a milder course and these individuals generally have normal lifespans. One third of the cases are new mutations. Sixty-five to seventy percent of patients demonstrate a deletion or in some cases a duplication. Assay: Direct analysis in males first via mutiplex PCR amplification of multiple dystrophin exons followed by Southern blot analysis with cDNA probes if indicated. Carrier detection in females by dosage analysis for deletions or by linkage analysis using intragenic and flanking polymorphic markers. Utility: Confirmation of diagnosis, carrier detection, prenatal diagnosis CPT codes: For PCR: 83890, 83894, 83901, 83912 for Southern: 83891, 83892, 83896, 83897, 83912 For linkage: 83891, 83892, 83894, 83896, 83897, 83898, 83912 Turnaround: PCR 1 week, Southern 3 weeks, Linkage 4 weeks Familial Adenomatous Polyposis (FAP) Background: Familial adenomatous polyposis (FAP) is an autosomal dominant disorder which typically presents with colorectal cancer in early adult life secondary to extensive adenomatous polyps of the colon. Polyps also develop in the upper gastrointestinal tract and malignancies may occur in other sites including the brain and the thyroid. Helpful diagnostic features include pigmented retinal lesions known as congenital hypertrophy of the retinal pigment, jaw cysts, sebaceous cysts, and osteomata. Screening for a mutation in the coding region of the APC gene is offered to individuals who have multiple colonic polyps. A pedigree demonstrating all cancers in the family must be submitted with the sample. Family members of individuals in whom a mutation has been identified can be studied to determine if they inherited the familial mutation. Assay: Conformation Sensitive Gel Electrophoresis/ Sequencing (analysis of PCR products for the presence of heteroduplexes followed by sequencing to detect differences from the consensus sequence of APC). Utility: To identify individuals at very high risk of developing colon cancer so that they can be targeted for aggressive prevention programs University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 25 and to reassure individuals in families with a known mutation that they are not at any higher risk than individuals in the general population (1-25 lifetime risk). CPT codes: Screening: 83891, 83898, 83903, 83904, 83912 Familial Mutation: 83890, 83894, 83898, 83903, 80904, 83912 Turn around: 8 weeks for screening, 2 weeks for familial mutation Fragile X Syndrome Background: This is the most common cause of mental retardation in males and also a common cause of learning disabilities and emotional dysfunction in males and females. Carriers can be mentally retarded or normal. The disorder becomes clinically manifest when the mutation (an expansion of a CGG nucleotide repeat) reaches a threshold of 600bp. Assay: Direct mutation analysis by Southern blot. Utility: Confirmation of clinical diagnosis, carrier detection, prenatal diagnosis CPT codes: 83891, 83892, 83896, 83897, 83912 Turnaround: 2 weeks Friedreich Ataxia (FRDA) Background: Friedreich Ataxia is the most common hereditary ataxia with an estimated prevalence of 1 in 50,000 and a carrier frequency of 1 in 100 in the European population. It is an autosomal recessive disorder characterized by progressive gait and limb ataxia, lack of tendon reflexes in the legs, loss of position sense, dysarthria, and pyramidal weakness of the legs. Hypertrophic cardiomyopathy is found in almost all patients and diabetes mellitus is seen in 10% of the patients. The age of onset is usually around puberty, almost always before 25 years with a slow progression of the disease. The disease is due to expansion of an unstable GAA trinucleotide repeat in intron 1 of the frataxin gene. Assay: Direct mutation detection by long range PCR and/or Southern blot, sequence analysis for point mutation if indicated. Estimated 95% detection rate. Utility: Confirmation of diagnosis in a symptomatic individual, predictive testing for asymptomatic individuals with a positive family history, carrier testing, and prenatal testing. CPT Codes: 83891, 83892, 83894, 83896, 83897, 83898, 83904, 83912 Turnaround: 3 weeks Hemophilia A Background: This is the most common severe bleeding disorder which is inherited as a classic Xlinked recessive. In half of all cases of severe Hemophilia A (factor VIII <1%) an inversion is detectable. In those cases where the inversion is not present or if the Factor VIII level is not <1%, direct mutation analysis and linkage analysis are available. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 26 Assay: Direct detection of the inversions by RT-PCR and direct mutation analysis by CSGE [Conformation Sensitive Gel Electrophoresis / Sequencing. PCR amplified products, including all coding exons and the exon-intron boundaries, are analyzed by electrophoresis in a novel polyacrylamide gel system for the presence of aberrantly migrating DNA fragments indicating sequence differences from the consensus sequence of Factor 8. Utility: Diagnostic confirmation, carrier detection, and prenatal diagnosis CPT codes: Inversion detection: 83891, 83892, 83896, 83897, 83912 Direct mutation analysis: 83891, 83898, 83903, 83904, 83912 Known mutation: 83890, 83894, 83898, 83903, 83904, 83912 Linkage: 83890, 83892, 83894, 83896, 83898, 83912 Turnaround: inversion testing 2 weeks, known mutation 2 weeks, direct mutation screen 2-15 weeks, linkage 3-4 weeks Hereditary Papillary Renal Carcinoma (HPRC) Background: Hereditary papillary renal carcinoma is a recently recognized form of inherited kidney cancer due to a mutation in the MET protooncogerne characterized by a predisposition to develop multiple bilateral papillary renal tumors. The pattern of inheritance is consistent with autosomal dominant transmission with reduced penetrance. Assay: PCR amplification, mutation scan by CSGE and sequence analysis of abnormal bands for those four exons where all mutations discovered to date were found. Utility: Early diagnosis so that increased surveillance could detect tumors early. CPT codes: 83890, 83894, 83898, 83903, 83904, 83912 Turnaround: 4 weeks HNPCC-Mutation Analysis of MSH2 and MLH1 (HNPCC/Muir Torre Syndrome) Background: Colorectal cancer (CRC) occurs in an estimated 130,000 patients in the United States each year. Five to ten percent of all CRC cases are thought to be inherited and the remainder is sporadic. The most common form of hereditary CRC is HNPCC (hereditary nonpolyposis colon cancer). Approximately 70% of families that meet the established Amsterdam and Bethesda diagnostic criteria will have a mutation in one of the 4 genes, which have been identified so far in HNPCC. Over 90% of these will have a mutation in MSH2 or MLH1. Eligibility: Patients who have or have had CRC and who meet the Amsterdam or Bethesda criteria are eligible for testing. Also isolated patients with CRC and sebaceous skin cancer may carry mutations in MSH2 and are eligible for testing. The Amsterdam criteria are defined as 1) at University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 27 least two successive generations should be affected with CRC; 2) diagnosis of one affected individual before the age of 50; 3) histologically verified CRC in at least three relatives, one of whom is a first-degree relative of the other two. The Bethesda criteria consist of modifications of the Amsterdam criteria and include 1) very small families which cannot be expanded further with two documented cases of CRC or families with 2 first degree relatives with CRC and the presence of a third relative with an early-onset unusual neoplasm or endometrial cancer; 2) age 55 instead of 50 as an upper limit of early-onset cancer; and 3) verification of cancer may be death certificates or clinical charts. Assay: Conformation Sensitive Gel Electrophoresis (CSGE) Analysis of PCR products for the presence of heteroduplexes followed by sequencing to detect differences from the consensus sequence of MSH2 and MLH1. Utility: To identify individuals at high risk of developing CRC, endometrial and other cancers so that they can be targeted for regular screening by colonoscopy and endometrial biopsy and to reassure other individuals in families with known mutations that they are not at any higher lifetime risk than individuals in the general population (6%). CPT codes: Screening-83891, 83898, 83903, 83904, 83912 Familial Mutation-83890, 83894, 83898, 80903, 83904, 83912 Turnaround: 8-12 weeks for screening, 2 weeks for familial mutation Huntington’s Disease (HD) Background: A severe autosomal dominant neurodegenerative disorder causing progressive choreiform movements and often dementia in mid-adulthood. The molecular defect is expansion of a CAG trinucleotide repeat. Assay: Direct detection by PCR analysis and gene scan of the number of CAG repeats in the Huntington gene. The primers used do not include an adjacent polymorphic region. Utility: Diagnostic confirmation, prenatal diagnosis, presymptomatic testing (presymptomatic testing is only available through the Neurogenetic Center Presymptomatic Huntington’s Disease Program or by special arrangement-talk with the genetic counselor) CPT codes: 83890, 83894, 83898, 83912 Turnaround: 2 weeks Li-Fraumeni Syndrome (P53) Background: This is an autosomal dominant cancer predisposition syndrome which predisposes to multiple forms of cancer (breast cancer, soft tissue sarcoma, leukemia, melanoma, osteosarcoma, colon cancer, pancreatic cancer, esophageal cancer, brain tumors and University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 28 others). The syndrome is due to germ line mutations in the p53 gene. This disorder is defined as: a proband with a sarcoma under 45 years of age and a first degree relative with any cancer under 45 years of age and a third family member who is a first- or seconddegree relative with cancer under 45 years of age or a sarcoma at any age. Eligibility: Patients who have or have had one of the index cancers and whose family history meets the Li-Fraumeni criteria are eligible for screening. Individuals from families where a mutation has already been identified are eligible for specific mutation testing. Assay: Conformation Sensitive Gel Electrophoresis (CSGE). Heteroduplex analysis of PCR products of genomic DNA of the p53 in a novel polyacrylamide gel for the presence of heteroduplexes. Differences between the 2 alleles of an individual as small as a single base substitution, insertion or deletion are detected as 2 bands. Sequencing follows the preliminary screening to identify the exact nature of the alteration. 1. C.J.Williams, M.Rock, E.Considene, S.McCarron, P.Gow, R.Ladda, D.Mclain, V.Michels, W.Murphy, D.J.Prockop and A.Ganguly. (1995) Hum. Mol. Genet. 4, 309-312. 2. A.Ganguly, M.J. Rock and D.J.Prockop (1993) Proc. Natl. Acad. Sci . U.S.A. 90, 10325-10329. 3. A. Ganguly and C. Williams. (1997) Hum.Mutat. 9, 339-343. Utility: Diagnostic confirmation, carrier detection, and prenatal diagnosis CPT codes: Direct mutation analysis: 83890, 83894, 83898, 83898-76, 83912 Familial Mutation-83890, 83898, 83898-76, 83912 Turnaround: 10-14 weeks Multiple Endocrine Neoplasia (MEN2A) Background: MEN2A is an autosomal dominantly inherited cancer syndrome characterized by medullary thyroid carcinoma, pheochromocytoma, and parathyroid hyperplasia. In 97% of MEN2A patients, the mutation involves one of five cysteine residues in the extracellular domain of the RET proto-oncogene. Assay: PCR amplification of genomic DNA, restriction enzyme digestion, and sequencing of exons 10, 11, 13 & 14. Utility: Testing is useful to confirm a clinical diagnosis or to identify presymptomatic carriers in a family. CPT codes: 83890, 83894, 83898, 83904, 83912 Turnaround: 2 weeks Multiple Endocrine Neoplasia 2B (MEN2B) Background: MEN2B is an autosomal dominantly inherited cancer syndrome characterized by bilateral medullary thyroid carcinoma, pheochromocytoma, diffuse ganglioneuromas of the University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 29 intestinal tract, mucosal neuromas, and skeletal abnormalities. Symptoms of MEN2B are evident within the first decade of life and progresses rapidly. In greater than 95% of cases, there is a mutation at a single position within the extracellular ligand-binding domain of the RET proto-oncogene. Assay: PCR amplification of genomic DNA and restriction enzyme digestion. Utility: Confirm clinical diagnosis; identify presymptomatic carriers prior to prophylactic thyroidectomy. CPT Codes: 83890, 83892, 83898, 83912 Turnaround: 1 week Retinoblastoma Background: Retinoblastoma (RB) is a malignant tumor of the developing retina that occurs in children, usually before the age of five years. Retinoblastoma may be unilateral or bilateral. About 60% of patients have unilateral RB with a mean age of diagnosis of 24 months; about 40% have bilateral RB with a mean age of diagnosis of 15 months. Mutations in the RB gene (chromosomal location 13q14) predispose individuals to the disease and an increased risk of developing other RB-related (non-ocular) tumors. Assay: Direct mutation analysis by CSGE [Conformation Sensitive Gel Electrophoresis] and sequencing. PCR amplified products, including all coding exons and the exon-intron boundaries, are analyzed by electrophoresis in a novel polyacrylamide gel system for the presence of aberrantly migrating DNA fragments indicating sequence differences from the consensus sequence of RB1. Mutation analysis by CSGE of the RB gene can identify a germline mutation in about 70% of individuals with a hereditary predisposition to RB. The probability that an RB1 gene mutation will be detected in an index case depends upon whether the tumor is unilateral or bilateral and whether the family history is positive or negative. Utility: Diagnostic confirmation, carrier detection and prenatal diagnosis CPT codes: 83891, 83898, 83898-76 (x8), 83904, 8390476 (x8), 83910 Sensitivity: If the patient has a mutation in the coding sequencing of the RB1 gene, the testing will detect the mutation ~70% of the time. Mutations in non-coding sequences, insertions, deletions or rearrangements will not be detected by sequencing. Sickle Cell Anemia (Hb SS or SC) Background: Autosomal recessive disorder of red blood cells which causes them to become distorted leading to blockage of capillaries, pain and damage to various organ systems. Assay: Direct mutation detection by PCR Utility: Confirmation of carrier or affected status and prenatal diagnosis University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 30 CPT codes: 83890, 83892, 83894, 83894, 83898, 83912 Turnaround: 1 week von-Hippel Lindau (VHL) Background: von-Hippel Lindau disease is a potentially lethal autosomal dominant disorder predisposing to benign and malignant tumors of the brain, spine, retina, pancreas and kidneys. Sensitivity is 93%. Assay: Rearrangement by Southern and/or direct mutation detection by sequence Utility: Early diagnosis so that increased surveillance could detect tumors early and prevent death GENETIC DIAGNOSTIC LABORATORY 25 from hemangioblastomas and renal cell carcinomas, prenatal diagnosis CPT codes: Southern blot: 83891, 83892, 83896, 83897, 83912 Sequence:83891, 83894, 83898, , 83903, 83904, 83912 Known point mutation: 83890, 83892, 83894, 83898, 83904, 83912 Turnaround: 2 weeks for Southern, 2 weeks for sequence, 2 weeks for familial mutation U N I V E R S I T Y O F P E N N S Y LVA N I A SCHOOLOFMEDICINE TRANSGENIC & CHIMERIC MOUSE FACILITY Nancy E. Cooke, M.D., Facility Co-Director 215/898-4425 necooke@mail.med.upenn.edu Stephen A. Liebhaber, M.D., Facility Co-Director 215/898-7834 liebhabe@mail.med.upenn.edu Jean Richa, Ph.D., Technical Director 215/898-6064; 215/573-3023 jricha@mail.med.upenn.edu Pei Fu He, Ph.D., Research Specialist 215/573-3023 peifu@mail.med.upenn.edu Kathleen A. Thompson, Research Specialist 215/573-3023 kthompso@mail.med.upenn.edu University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 31 Location: 32 Clinical Research Building Mailing Address: 502 Clinical Research Building 415 Curie Boulevard Philadelphia, PA 19104-6145 Telephone: 215/573-3023 Fax: 215/573-5892 Internet: http://www.med.upenn.edu/tcmf The Transgenic and Chimeric Mouse Facility provides a centralized service of transgenic mouse production to the Penn community. It offers services such as DNA injection into fertilized oocytes, ES cell injection into blastocysts, line rederivation, and embryo cryopreservation, along with the care and housing of the mice up to the stage where they will be transferred to the participating investigator. The facility is fully equipped to carry out these procedures. The facility is located behind a microbiologic barrier where admittance is restricted and all food, water, cages, and bedding are autoclaved before use. 2004 GENETICS CORE FACILITIES CATALOG 28 Introduction The Transgenic Mouse System Integration of genes into the mouse genome can serve as an experimental system to study normal as well as altered gene expression. Such studies can also serve to identify genes essential for normal development and generate animal models for human diseases. Direct microinjection of DNA into the male pronucleus of a mouse zygote has been the method most extensively used in the production of transgenic mice. If the foreign DNA integrates into the mouse chromosomal DNA at the onecell stage, the transgenic animal will contain the injected DNA in every cell, including those of the germ line. If, however, integration occurs later, the resulting animals will be mosaic for the presence of the injected DNA. The number of copies of integrated DNA can vary from one to several hundred, arranged primarily in tandem head to tail arrays. The production of chimeric mice by introducing Embryonic Stem (ES) cells into early embryos is an alternative approach to generating transgenic mice. This technique utilizes the tissue culture system to modify and select ES cells that have received an exogenous DNA of interest. Once derived and characterized, ES cell clones may be transferred into 4-day-old mouse embryos where they can differentiate into any adult tissue type. The choice of mouse strain is of great importance: donor embryos that would generate ES cells and host embryos that would receive transferred cells are obtained from different strains of mice carrying different eye or coat pigmentation. This facilitates the visual distinction between cell populations in the adult “chimeric mouse”. The Facility The purpose of the Transgenic & Chimeric Mouse Facility is to provide a centralized service to efficiently produce transgenic mice for basic research. This should result in reduction in effort and cost to participating investigators. The facility is located on the basement level of the Clinical Research Building. This facility consists of an injection room and several animal rooms. The injection room is fully equipped to carry out the entire procedure of making transgenic mice. The animal rooms provide housing and breeding space for the mice involved in the transgenic projects. The facility uses sterile food and water as well as University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 32 autoclaved cages and bedding; all cages are of the microisolator type to limit the spread of colony infection. The entire facility is located behind a microbiologic barrier where admittance is strictly limited and all personnel must wear sterile coveralls, gloves, hats, masks, and boots. Personnel Nancy E. Cooke, M.D., is the Facilitity Director and Stephen A. Liebhaber, M.D., is the Facility Co-Director. Jean Richa, Ph.D., is the Technical Director in charge of the production of transgenic mice. Dr. Richa will maintain direct contact with investigators using facility, coordinate ULAR interactions and facility maintenance, and be directly responsible for scientific consultations relevant to faiclity use. Pei-Fu He, Ph.D. and Kathleen A. Thompson, Research Specialists, assist in various aspects of the facility’s services. What the Facility will Provide DNA Injection Upon receipt of the DNA construct, proven by documentation to be of high quality and suitable for microinjection (see below), we will proceed to inject a minimum of 150 fertilized eggs. After overnight incubation, normal 2-cell embryos will be transferred into foster females and their pregnancy will be monitored (the gestation period of the mouse is 19-20 days). Two weeks after birth of potential transgenic pups, the project scientist will be notified of the number of pups. The facility will carry out tail clipping and animal tagging prior to weaning. Tail biopsies will be conveyed to the project scientist for analysis by Southern blotting, PCR, or both. At this point, the clipped and tagged pups will be transferred to the care of the individual investigator who will be responsible for subsequent maintenance. Since cage costs are high and storage space is limited, investigators should make certain that DNA screening procedures are working prior to initiating transgenic work with the facility. Investigators must report the results of screening with documentations (e.g., gel photo, etc.) to the Technical Director. This information is necessary for facility personnel to maintain and improve their efficiency. If these studies (with adequate controls) fail to demonstrate the existence of at least one transgenic animal, we will reinject at least 80 eggs at no additional cost to the investigator. If, however, no transgenic animal is detected after the second trial, a meeting will be held between the scientist, the Facility CoDirectors, and the Technical Director to discuss the DNA construct, the tail biopsy analysis, and further plans to be agreed upon. On average, 230 injected eggs should yield at least one (and usually more) transgenic mouse. It is well understood that many factors can affect the production efficiency of transgenic mice. Such factors include: _ Number of eggs surviving the injection and developing to 2-cell stage; this will determine the number of transfers into foster females. It is important to thave a DNA preparation pure of any contaminant and at the appropriate concentration so that toxic effects to the eggs can be avoided. _ Successful pregnancies: although every transfer promises a certain number of pups, that number may vary greatly due to embryo death in utero. This University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 33 lethality may be closely associated with the type of DNA construct used. Transient transgenics For investigators who wish to examine DNA integration and expression in transgenic embryos, we can provide similar DNA injection schedule for a fee of $855.00. We will inject 150 eggs, transfer the developing embryos into foster females, and arrange with the project scientist to receive the pregnant females on the asssigned date. We would still need to receive a report on the number of embryos harvested from each female, and the frequency of transgenics among them. ES cell injection Through this service, the facility will generate chimeric mice by injecting Embryonic Stem (ES) cells containing site specific genomic alterations (knockouts) into normal blastocysts. ES cells will be injected into a minimum of 50 blastocysts of C57BL/6J. When the blastocysts have reexpanded, they will be transferred into the uterine horns of pseudopregnant females and allowed to go to term. Upon weaning 3 weeks after birth, chimeric mice are separated and then released to the care of the investigator. Due to the fact that this approach is time-consuming, it is recommended that certain guidelines be followed in order to minimize loss of time and reduce risk factors. These guidelines include: _ Before initiating the injection of the recombinant cell line, normal non-recombinant ES cells should be tested for their chimera-producing potential. _ A mycoplasma test of the recombinant line is absolutely necessary since the presence of such contamination has been associated with loss of pregnancy and reduced litter size. _ On the assigned day, the freshly harvested ES cell sample (free of feeder cells) will be delivered to Dr. Jean Richa to proceed with this injection. _ A photocopy of the recombinant ES cell line screening results (Southern blot or PCR) should be submitted along with the chimeric mice request form (see page 13). Embryo cryopreservation Freezing mouse embryos is a convenient way to preserve in important mouse line for possible future use or to protect this line against loss due to infection, lab accident, etc. Embryo freezing could also result in substantial reduction in housing cost and cage use This newly added service is now available to interested investigators. The service can be designed to suit the needs of each investigator. However, a general scheme is outlined below. A set of five young male mice will be needed to generate enough embryos for each freezing set. The male founders will be subjected to a waiting period in the containment area, as required by ULAR, prior to admission into the barrier colony. A minimum of 300 healthy embryos will be frozen per transgenic/knockout line that is hemizygous for the transgene. In the case where the mouse line is homozygous for the transgene, 150 embryos will suffice. Several weeks after the completion of the freezing process, a few embryo samples will be removed and processed through thawing and monitoring development to assess University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 34 their viability and ability to develop to term, Once these tests are completed, the frozen embryos will be submitted to the care of the investigator for proper storage. High efficiency of embryo freezing will depend primarily on the reproductive level of the breeders since low mating frequency and low percentage of fertilization will lead to a prolonged time-consuming process. Mouse line rederivation This process involves the surgical transfer of embryos into pathogen-free recipient females, where they can develop to term. Rederivation is the most effective method to rid a colony of pathogenic infections. It is also recommended when genetically altered animals are acquired from outside institutions that do not fall into the category of approved vendors. With the proper procedure, mouse lines can be saved or transferred safely across distant locations. Due to the diverse environmental conditions among the animal facilities located across campus, the structure of this service will be tailored to fit each and every facility receiving the rederived line. As a consequence, the fees may vary accordingly. Basically, a total of 50 embryos generated from the designated mouse line will be transferred into recipient pathogen-free females that would carry them to term in a housing location assigned by ULAR. The mice born from such transfers should be of sufficient number to rejuvenate or establish the line. Investigator’s Responsibilites For DNA injection In general terms, the investigator must prepare the DNA to be injected and assume responsibility for screening and care of the weaned animals. Specifically, in order to provide you with this service, it is requested that you: 1) Contact Jean Richa (215/898-6064) to discuss the DNA construct and the schedule of injections. 2) Provide the facility with a high-quality DNA preparation of your construct with documentation of its purity. A collection of four tubes appropriately labeled and each containing 25 µl of DNA at 5 ng/µl concentration would suffice. (The recommended protocol for DNA preparation can be found in the Appendix.) 3) Fill out and sign a service request form, including evidence of approval of the IACUC and IBC committees for the project. Unless you have been doing transgenic mouse work, this will require a new submission to both committees. A sample of such a form is on page 11 of this manual. Feel free to duplicate it as you need it. 4) Provide a timely and well-controlled analysis of the tail biopsies so that transgenic animals can be identified appropriately and your cage costs can be minimized. 5) Undertake the care of the potential transgenic pups submitted to the project scientist along with the tail biopsies. These mice can no longer reenter the barrier colony. It is the responsibility of the individual investigator to arrange for animal housing. For ES cell injection The investigator is responsible for maintaining the recombinant ES clone under optimal conditions that would inhibit their differentiation. Once the chimeric animals are weaned, it is the responsibility of the investigator to breed them for germ line transmission testing. It is recommended University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 35 that you: 1) Contact Jean Richa (215/898-6064) to arrange for testing the non-recombinant ES cell line and discuss the schedule of injection. 2) Fill out and sign a service request form, including evidence of approval of the IACUC and IBC committees for the project. Unless you have been involved with chimeric mouse work, this will require a new submission to both committees. A sample of such form is on page 13 of this manual. Feel free to duplicate it as you need it. 3) Upon notification of a prearranged schedule, provide the facility with a freshly harvested sample of ES cells along with an aliquot of HEPES-buffered ES cell culture medium (5 ml). The cell sample should be free of feeder cells. Additional information on maintaining and harvesting ES cells could be found in Teratocarcinomas and Embryonic Stem Cells (E.J. Robertson, Ed.), IRL Press, Oxford, 1987, as well as in Meth. Enz. v. 225, 1993. 4) Undertake the care of the weaned 3-week-old chimeric mice submitted to the project scientist. These mice can no longer reenter the barrier colony. It is the responsibility of the principal investigator to arrange for animal housing. 5) Relay to the facility any information about germ line transmission once such information becomes available. For Embryo Cryopreservation The investigator should contact Jean Richa (215/898-6064) to discuss the plans for the service. These would include the availability of five breeding males, the possibility of admitting the animals into the barrier facility, the number of mouse lines involved, and the service fees. For Line Rederivation The investigator should contact Jean Richa (215/898-6064) to discuss the plans for the service. These would include the availability and housing location of the line(s) to be rederived, as well as the service fee. The investigator should be responsible for the housing and care of the recipient females while they are in transition in the location assigned by ULAR. Costs We will charge a fee of $1,100.00 per DNA construct injected for the production of transgenic mice. The initiation of zygote injections depends upon receipt of an approved protocol and request form accompanied with properly labeled tubes of the DNA construct and documentation of its purity. Currently, we harvest fertilized eggs from superovulated B6SJLF1/J females mated with B6SJLF1/J males (from JAX). We will attempt to accommodate requests for other mouse strains if required by specific experiments. Please justify these requirements in your protocol for review. Additional costs will be incurred. In the case of DNA injection for transient transgenics, the fee will be $855.00 per DNA construct. For ES cell injection, a fee of $1,100.00 will be charged per cell line injected for the production of chimeric mice. The initiation of blastocysts injections depends upon receipt of an approved protocol and request form, following a schedule prearranged with Dr. Jean Richa. The schedule University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 36 will be discussed after testing the ES cell line and approving the documentation of the recombination event in the assigned ES recombinant clone. Currently, we harvest blastocysts from C57BL/6J females mated with C57BL/6J males (from JAX). An initial, one time fee of $300.00 is charged for testing the non-recombinant ES cell line for its ability to contribute to chimerism. The full cost of the transgenic service (DNA/ES) is $3,780.00 per construct/cell line. The low fee of $1,100.00 for Medical School investigators, and members of the Cancer Center, Diabetes Center, and Digestive and Liver Diseases Center, is possible due to a substantial subsidy by those organizations. For all other Penn investigators, the cost of either service is $2,200.00 per construct/cell line. For embryo cryopreservation, the cost will depend primarily on the successful reproduction of the breeding males provided. A minimum estimate of $1,100.00 is projected, with the cost of the quarantine housing being the responsibility of the investigator. For line rederivation, the cost will depend primarily on the source location as well as the destination of the mouse line to be rederived. A minimum estimate of $550.00 is projected. Policies Committee Approval It is an Institutional requirement that every investigator demonstrate an updated approval by the Institutional Animal Care and Use Committee (IACUC) as well as the Institutional Biosafety Committee (IBC) for both DNA and ES cell injection projects prior to using the Transgenic & Chimeric Animal Facility. Scheduling It is our policy that no injection date will be assigned to any DNA construct until it is submitted along with a completed TRANSGENIC & CHIMERIC MOUSE FACILITY 31 service request form and the required documents attached. Confidentiality All information provided by the investigator concerning the research project are treated with the utmost confidentiality. No subject related material will be released to or shared with anyone without a written consent of the principal investigator. Support Letter For those investigators preparing to submit grant proposals to funding agencies, we will be happy to provide you with a letter stating our readiness to make transgenic and/or chimeric mice for your project. Acknowledgments We request that any scientific publications involving these transgenic animals acknowledge the service of the University of Pennsylvania Transgenic & Chimeric Mouse Facility in the Materials and Methods section. Submitting Results Results of the transgene screening should be reported back to the TCMF using the Transgene Screening Results form (see page 12). Failure to report results within 45 days of receiving the tail samples will result in the forfeiture of your right to a free re-injection in the event that only 0-2 founder mice can be detected. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 37 Housing Arrangement Tails samples and mice should be received at the same time by the investigator or the project scientist. We can no longer hold the mice while the samples are being screened for the presence of the transgene. The P.I.’s should prearrange for space availability in their assigned facility. Appendix Purification of DNA for Microinjection 1. Recombinant plasmid should be purified by CsCl gradient (for details see Molecular Cloning: A laboratory manual, 2nd ed. Sambrook, et al. 1990). The insert is then released with the appropriate restriction enzymes. It is critical to remove all vector sequences, as such sequences can significantly alter the expression of transgenes (mechanism unclear). 2. Separate the insert of interest from the vector on an agarose gel run in Tris/Acetate/EDTA (not Tris/ Borate/ EDTA) buffer. 3. Excise the gel slice containing the gene fragment of interest and electroelute the DNA fragment. 4. Recover the fragment by ethanol precipitation, and then pass it through an ion exchange column (e.g., Schleicher & Shuell Elutip columns) according to the manufacturer’s instructions. 5. Ethanol precipitate the DNA and redissolve it in injection buffer (10 mM Tris/0.1 mM EDTA pH 7.5). This buffer must be prepared with Milli-Q water to maintain zygote viability. We can provide this type of water upon request. 6. Estimate the DNA concentration by comparing the ethidium bromide staining of a sample run on an agarose gel next to a standard of known concentration such as • DNA cut with Hind III. 7. Adjust the DNA concentration to 5 ng/µl with injection buffer. Alternate Procedure 1. After separating the fragment on agarose gel run in TAE, excise the fragment of interest from the gel. 2. Process the gel fragment through Geneclean II kit (Bio 101, Inc.) or Qiaex gel extraction kit (Qiagen, Inc.) according to the manufacturer’s instructions. 3. Ethanol-precipitate the recovered DNA and resuspend in Elutip buffer 4. Pass through an Elutip Column (Schleicher & Shuell). 5. Ethanol precipitate the recovered DNA and resuspend in injection buffer (10 mM Tris/0.1 mM EDTA, pH 7.5 prepared with Milli-Q water). 6. Estimate the DNA concentration by comparing the ethidium bromide staining of a sample run on an agarose gel next to a standard of known concentration. 7. Adjust the DNA concentration to 5 ng/µl, using injection buffer. Notes _ For ethanol precipitation of a sample, add 1/10 volume of 3 M NaAc and 2-2.5 vol. ethanol. _ Use 1:100 dilution of EtBr (5 mg/ml) for gel staining. Visualize the DNA band with long-wave UV light to avoid DNA damage. _ It is strongly recommended that powder-free gloves be used when handling the DNA preparation. Powdered University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 38 gloves are most often associated with the presence of particulate matter that gets lodged at the tip of the injection needle and renders it useless. Protocol for Isolating High-Molecular-Weight DNA from Mouse Tails (From Manipulating The Mouse Embryo; Hogan et al. Cold Spring Harbor, 1986). Day 1 1. Cut 0.5-1.0 cm of tail and place in a 1.5-ml microfuge tube (tails can be frozen at -20 •C prior to extraction). These tail biopsies will be performed by the Core. 2. Add to the tube 0.7 ml of 50 mM Tris (pH 8.0), 100 mM EDTA, 0.5% SDS. Add 35 µl of a fresh 10 mg/ml solution of Proteinase K. 3. Incubate at 55 •C overnight on a gently rocking platform. Day 2 4. Next morning, remove tubes from 55 •C. Add 0.7 ml of phenol (equilibrated with Tris - pH 8.0). Close tube and shake vigorously for 3 minutes, so that phases mix completely. 5. Centrifuge in a microfuge for 3 minutes. Phases will separate. 6. Transfer upper aqueous phase to a fresh tube, being careful not to pick up phenol or material at interface. 7. Add 0.7 ml of phenol/chloroform (1:1), shake vigorously for 2 minutes and centrifuge for 2 minutes. 8. Again remove aqueous phase, avoiding interface, and transfer to a 1.5-ml tube. 9. Add 70 µl of 3 M sodium acetate / pH 6.0 (i.e., 1/10 volume), and 0.7 ml of 100% ethanol at room temperature. Shake to mix thoroughly. DNA should immediately form a stringy precipitate. Sodium acetate with a pH lower than 6.0 will cause the EDTA to precipitate. 10. Spin in microfuge for 30 seconds to pellet DNA. Remove and discard as much ethanol supernatant as possible. 11. Add 1 ml of 70% ethanol (room temperature) to tube, and vortex or shake vigorously to wash DNA. This step is essential to remove traces of SDS and phenol. 12. Centrifuge in microfuge for 1 minute at room temperature. Remove as much ethanol supernatant as possible. Dry DNA briefly in vacuo. 13. Add 0.1 ml of 10 mM Tris (pH 8.0)/1 mM EDTA to tube. Leave at room temperature overnight to dissolve. If necessary, DNA can be dissolved more quickly by heating for 5-10 minutes at 65 •C. The DNA should have an A260/A280 of > 1.7, and the concentration should be calculated using: 50 µg/ml = 1.0 OD260. Use 10 µg of each DNA preparation for Southern blot analysis. 14. DNA prepared in this manner will contain substantial amounts of RNA, but this does not interfere with restriction enzyme digestion or Southern blot analysis. When performing restriction enzyme digestion, add 5 µg of DNase-free RNase A to each sample along with the restriction enzyme. Digestion with certain restriction enzymes may also be aided by adding 4 mM spermidine to the reaction. 15. Make sure you include the appropriate positive and negative hybridization controls for your Southern University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 39 analysis. Preparation of BAC DNA for microinjection Courtesy of Mike Thornton from the laboratory of Dr. Mortimer Poncz, Professor of Pediatrics at the Children’s Hospital of Philadelphia 1. Recombinant BAC plasmids should be purified using preferably the QIAGEN Large Construct Kit. This will yield at least 100 µg of pure plasmid free from the Genomic DNA contamination usually associated with BAC preps. CsCl gradient can be used alternatively (for details see Molecular Cloning: A laboratory manual., 2nd ed. Sambrook, et al 1990) with the following guidelines: a) After adding Solution II and/or III invert very gently about 8-10 times to ensure no genomic DNA is sheared, b) During all steps throughout this protocol use sterile wide bore pipettes, and minimize the amount of pipetting. c) Use a needle of size 18 gauge or wider to remove your band after ultra-centrifugation d) when you remove your band be absolutely sure not to remove the genomic DNA band that typically appears above the plasmid band. Once you have precipitated your DNA, the insert is then released with the appropriate restriction enzymes. 2. Separate the insert of interest from the vector sequence on a Seaplaque GTG(low melting) agarose gel run in 0.5-1X TBE (Field Inversion Gel Electrophoresis-FIGE Vs Standard electrophoresis respectively). 3. Preparation of the gel Reagents: _Baxter’s Sterile Water _TRIS-Base _Filter unit and receiver _Boric Acid _1000 ml clean glass bottles _EDTA-4Na _Nalgene 1000 ml sterilization 3a. 108g of Tris, 55g of Boric acid and 80ml of 0.5M EDTA-4Na (prepared with Millipore Water) are mixed by shaking in a sterile Nalgene receiver with 1000ml of Baxter’s water to prepare 10X TBE. This solution is then passed though a 0.2µm Nalgene 1000 ml filter unit. 3b. 50-100ml of 10X TBE (above) and 900-950ml of Baxter’s water are mixed and passed directly through a second 1000 ml filter unit and the resulting 0.5X(FIGE) or 1X (standard) TBE solution is transferred to a very clean 1L glass bottle for autoclaving. 3c. 100ml of 0.5 or 1X TBE is then added using an autoclaved glass graduated cylinder into a 500ml autoclaved flask, containing no greater than 1 %(w/v) Seaplaque GTG agarose. After heating make sure the volume is still 100ml and add 0.1- 0.2µl of Ethidium bromide, 10mg/ml stock (50-100 kb insert or < 50 kb insert respectively) 3d. Gel trays, combs and Electrophoresis chambers are washed with antibacterial soap and rinsed in millipore water before pouring or running gels. Appropriate care should be taken to keep the gels, gel trays chambers, etc., as sterile as possible. Using autoclaved pieces of aluminum foil to cover and protect items helps. 3e. Run molecular weight markers on both sides of the gel and also run expendable sample lanes on each side of the gel. The center lanes will contain the “real sample University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 40 “ to be isolated. Avoid exposing the center lanes to any light normal or UV. The sample and marker lanes on the sides can be used to monitor the progress of the run. 4. Excise the appropriate sized insert band from the gel. Supplies: _UV Box _Sterile Aluminum foil strips 10.5 cm X 7cm _New- sterile unused Scalpels 4a. Clean the surface of the UV box with Sterile Millipore water and/or 70% ethanol, air dry or dry using sterilized paper towels. 4b. Place two sterile aluminum foil strips on top of each other and place on the UV box. 4c. Place your gel (as little light as necessary in the room should be present) on the aluminum strips such that the marker lanes and sample lanes on both sides can be exposed to the UV light whereas the center lanes are on top of the foil protected from the UV light below. 4d. Using the marker and sample lanes on both sides as a guide place a clean sterile ruler under the desired band and use a scalpel to cut across the gel under the band, place the ruler above the band, and repeat. Cut away any excess gel on the sides of the desired sample region. Cut the gel slice into 6-8 pieces of equal size and place all slices into a 50-ml conical tube. Add 45ml of Baxter’s EQ buffer (below) 5. Equilibrate the gel slices in Baxter’s EQ buffer in 2 exchanges over 1-2 hours. Remove all buffer using a vacuum and sterile Pasteur pipettes. Place1-2 gel slices into a single sterile Eppendorf tube (4-8 tubes total) and melt the gel slices at 68 C for 10 minutes then place tubes in a 42 C water bath for five minutes to equilibrate. Baxter’s Buffer: Final Stock 10mM Tris pH 7.5 ...... 5 ml of 1M Tris pH 7.5 250µM EDTA ............ 250 µl of 0.5M EDTA pH 8.0 100mM NaCl ............. 10 ml of 5M NaCl Baxter’s water ........... ~484 ml Stock solutions used to make above dilutions should only use Millipore water and is filter sterilized before use. Add all components directly into 500 ml Nalgene filter sterilization unit. Pour filtered buffer in to a very clean glass bottle and autoclave. 6. Digest the agarose slices Add 10 µl of beta agarase enzyme (New England Biolabs) to every 500µl of melted gel slice, mix (pipette with wide pore sterile tips) and incubate overnight at 42 C. The next day place tubes back at 68 C for 10 minutes and then 42 C for five minutes add 2µl more of beta agarase and incubate for an additional 1-2 hours. Centrifuge tubes for 10 minutes full speed to get rid of any remaining debris or undigested agarose. 7. Concentrate the insert DNA Carefully transfer your sample from each Eppendorf tube into a Millipore Ultrafree - 4ml Centrifugal filter, specifically the Biomax 50k size exclusion type (Catalog # UFV45010B) This filter needs to be prewashed with sterile water or Baxter’s EQ buffer to University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 41 get rid of the glycine preservative found on all new filters. Be careful not to transfer any undigested agarose or debris at the bottom over to the centrifugal filter. Centrifuge the sample for 5 minutes at 5001000g. Your initial volume should fall between 3-4ml before centrifugation and drop no lower than 500 µl after spinning. The final solution has the same initial salt concentration but increased DNA concentration. It is advisable to keep the flow-thru as a precaution. You can optionally add 3ml of T.E. buffer (10mM Tris pH 7.5 0.1 mM EDTA-4Na pH 8.0) prepared with Sigma Water for embryo Transfer (Catalog #W-1503) This will further dilute any proteins and salts remaining. Centrifuge again at 500g for 5 minutes or until the volume reaches 500 µl. Let the sample sit for one hour, than pipette gently and transfer to an Eppendorf tube. Filter the sample by placing it into a 1 cc syringe and passing it though a 0.2µM Acrodisc filter (Gelman Sciences Ann Arbor Michigan) This filter is specially designed to not stick to DNA. 7a. Drop dialyze sample Place a Millipore 0.025µm VS filter (catalog# VSWP 025 00) on to 10ml of T.E. buffer (10mM Tris pH 7.5 0.1 mM EDTA-4Na pH 8.0) prepared with Sigma Water for embryo Transfer(Catalog #W-1503). Place a 50 µl drop of your sample in the center of the filter and leave overnight. The next day remove the DNA drop and place in an Eppendorf tube. Determine the concentration of the sample and dilute in filtered T.E., to reach a final concentration of about 2-3ng/µl. Microinject the DNA within 1-2 days Optional: To get rid of excess ethidium and for longterm storage use protocol below. 7b. Ethanol precipitate your sample After acrodisc filtering (step 6 above) - Add 1/10th the volume of sample of 3M NaOAc(autoclaved and filter sterilized( 0.2µM) and prepared with Millipore water) then add 2X volume filtered( 0.2µM ) 98% Ethanol place at -20 C until ready to use. This storage is good for several months. Centrifuge DNA and wash pellet in filtered 70 % ethanol, dry and add 200 µl of filtered (0.2µM) T.E. buffer(10mM Tris pH 7.5 0.1 mM EDTA4Na pH 8.0) prepared with Sigma Water for embryo Transfer. Drop dialyze the sample as above, and determine the concentration of the sample and dilute in filtered T.E., to reach a final concentration of about 2-3ng/µl. Microinject the DNA within 1-2 days Preparation of HNF3• YAC DNA for Microinjection Courtesy of Newman Sund and Dr. Klaus H. Kaestner, Department of Genetics, University of Pennsylvania School of Medicine For additional reference: Schedl, A. et al. (1993) Nucleic Acids Research. 21: 4783-4787. 1. Equilibrate 8 yeast plugs (Large Scale Yeast Plug Prep) 1x 10', 1x 30', 1x 1hr in 50 ml/wash of 0.40x TBE (BioRad 10xTBE #161-0733). Equilibrate “PFGE markerI-Lambda ladder”(BMB #1378961) at this time as well. It is recommended that you make 3.0 liters of 0.40xTBE for plug washes, gel and running buffer. 2. Load plugs into 4 cm. slot of 1.0 % BioRad PF-certified agarose gel in 0.40xTBE. Make a 200 ml gel using the preparative comb. Plugs are positioned vertically, University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 42 tightly next to each other, and flanked by • ladder. Seal the plugs in the gel with 1.5% SeaPlaque Low Melting Point (LMP) agarose. Note _ The original protocol uses a 1.0% SeaPlaque (FMC) LMP agarose PF gel. Non-LMP agarose is recommended for use because the 1% LMP agarose gel is fragile and difficult to work with in steps 5 - 7. When excising the YAC DNA in step 8 using the nonLMP agarose, make sure not to excise the 1% gel with the 4% gel. The non-LMP agarose will not melt at 68 •C in step 10. Using the non-LMP agarose may affect the migration of the YAC DNA into the 4% LMP gel and affect the yield. 3. Run gel as optimized previously. For HNF3• YAC: 515 s. switch, 20 hr. run, 6V/cm, 120 • angle, 14 •C, 2.4 L of 0.40xTBE. You want the current to run 120-145 amps during the run, closer to 120A at the beginning of the run. 4. After the run, do NOT stain the entire gel. Instead, cut off both sides of the gel (• ladder plus half of one lane each - see Figure 1) and stain the marker gel pieces in 0.5 µg/ml Ethidium Bromide (EtBr) (20 µL 10 mg/mL EtBr stock in 400 mL dH20) for 30 min. rocking. While marker gel pieces are staining, soak rest of the unstained gel in 1 x TAE buffer (Maniatis). It is recommended that 4L of 1 x TAE is made for all the washes and the running buffer. Observe marker gel pieces under UV and take picture of gel with ruler aligned with the top of the gel. Note the position of the YAC band as well as two reference chromosomes. 5. Now excise the YAC band and the two individual reference chromosomes from the unstained, pulse field gel (cut out ±2.5mm). Stain the rest of the gel to confirm that you cut out the correct band. 6. Equilibrate gel slices in 1 x TAE buffer 1x 1hr, 1x 2hr rocking very gently. 7. Position gel slices in a minigel tray with the YAC slice in the middle (see Figure 2). The gel slices should be positioned lengthwise, such that the DNA will migrate at a 90 • angle to the original PFGE run. Seal the sides of the gel pieces onto the tray with 4% LMP-agarose (NuSieve GTG, FMC) in 1x TAE to keep the gel pieces precisely positioned. Prepare the 4% LMP gel beforehand and keep at 48 •C. LMP agarose foams and boils over easily so watch it carefully as it heats. Cast 4% LMP-agarose in 1 x TAE around the slices, barely covering them (~180-200 mL). Make sure that the surface is level when casting and running the gel. Keep some tape on tray ends to prevent the gel from sliding off during the run. Electrophorese for 16 hours at 60V (2.2V/cm) while recirculating the buffer. 8. Excise one entire reference chromosome plus at least 3 cm into the 4% gel where the DNA has migrated into and stain the gel piece in EtBr, as above. If there is still a significant amount of DNA in the 1% gel, electrophorese longer at 110V (4V/cm) for an additional 1-2hr and stain the other reference chromosome. Estimate the position of the YAC based on the position/location of the reference chromosome DNA and cut out the minimal YAC band. Place into tared µfuge tube and determine volume of the block. 1 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 43 mg=1µL. If necessary, cut the agarose block into smaller pieces such that each block is approximately 100-200 mg. This aids melting in step 10 and potentially concentrates the DNA in some aliquots. Stain the rest of the YAC slice to confirm you cut it out. (see Figure 3) 9. Equilibrate gel block 1x 10', 2x 1hr. in 15 ml/wash of 1 x TAE, 100 mM NaCl, 30 µM Spermine, 70 µM Spermidine. 10. Melt agarose block for 20 min at 68 •C, stirring the solution after 10 minutes with a sterile pipette tip. Cut the gel block into smaller pieces to aid melting. Do not incubate agarose at temperatures greater than 70 •C, as it will denature the DNA. Once melted, transfer tube immediately into 42 •C water bath and equilibrate the FIGURE 1 FIGURE 2 FIGURE 3 molten agarose for 10 minutes at 42 • C. GELase loses activity at temperatures greater than 45 •C. Add 5 units (1U/µL) GELase (Epicentre) per 100 mg agarose and incubate at 42 •C for 2 hours. While in heat block, stir molten agarose a couple of times with pipette to ensure even distribution of GELase. After 2 hours, place test tube on ice for 10 minute to test if digest is complete. If gel reforms, repeat step 10 with more GELase. 11. Pulse briefly at 3000 rpm for 10 sec. Dialyze against YAC micro-injection buffer for 3 hours. For this, place a Dialysis Membrane (0.05 microns, type VM, Millipore) on top of 50 ml of YAC micro-injection buffer in a Petri-dish. Pipette YAC DNA solution carefully on top of the membrane using a cut-off blue tip, dialyze for 3 hours, and transfer YAC DNA to a fresh µfuge tube. Save a 1 ml aliquot of YAC micro-injection buffer for dilution of YAC DNA in step 13. 12. Estimate YAC DNA amount by PFGE (load 20 µL) against known standards (10, 20, 40, and 80 ng). Include positive control and • marker as well. Can stop run at 18.5 hours to see plasmid stds. Notes for handling YAC DNA _ Always tap mfuge tube to mix purified YAC DNA prior to taking off an aliquot. _ Never vortex or pipette up and down to mix. This will shear the linear YAC DNA. _ Use a cut-off pipette tip when you need to take an aliquot of YAC DNA. _ Store YAC DNA at 4 •C until ready for use. 13. Dilute YAC DNA to 0.3-2.0 ng/µL. You have to dilute at least 2:1 with YAC micro-injection buffer: YAC DNA for micro-injection. Therefore, the YAC DNA must be at minimum 0.9 ng/µL when quantitated. YAC micro-injection buffer (10 mM Tris/Cl pH 7.5, 0.1mM EDTA pH 8.0, 100 mM NaCl, 30 µM Spermine, 70 µM Spermidine) For 200 mL 1M Tris pH 7.5 2.0 mL 5M NaCl 4.0 mL 0.5M EDTA 40 µL 0.3M Spermine (1:10,000 diln) 20 µl 0.7M Spermidine 20 µL MQ ddH20 194 mL 1xTAE, 100 mM NaCl, 30 µM Spermine, 70 µM Spermidine University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 44 For 200 mL 50xTAE 4.0 mL 5M NaCl 4.0 mL 0.3M Spermine 20 µL 0.7M Spermidine 20 µL MQ dd H20 192 mL 0.3M Spermine (348 g/mol) For 1 ml: 0.104g in 1.0 ml ddH20 0.7M Spermidine (255 g/mol) For 1 ml: 0.179 g in 1.0 ml ddH20 Large Scale Preparation of Yeast Plugs 1. Inoculate 100 ml medium (YPAD or selective medium) with 0.2 to 1 ml of a stationary culture. Shake at 30 •C for 1 to 2 days. Density should be at least 5 x 107 cells/ml. 2. Pellet cells in 50 ml Falcon tubes at 3000 rpm for 5 min. 3. Resuspend pellets in 40 ml (total) 50 mM EDTA, pH 8. Spin as above 4. Weigh a Falcon tube. Resuspend cells in Solution 1 and pellet in Falcon tube. 5. Determine the weight of the pellet. Assuming 1g = 1ml, you now know the volume of the pellet. Add 0.5 volume Enzyme Solution and warm to 37 •C. 6. Now add equal volume of pre-warmed (40 •C) Agarose Solution. Suspend the cells using a cut-off blue tip. Pipett into 8- microliter block forms. 7. Place on ice for 10 min. Gently remove plugs and incubate for 2 hours at 37 •C in 5 ml per plug of Solution 2. 8. Transfer to LDS at 37 •C. After one hour, transfer to fresh LDS and incubate (gentle shaking) at 37 •C overnight. 9. Next day, wash plugs for two hours in 20% NDS at room temp. Store plugs indefinitely at RT. 10. Before use, plugs have to be equilibrated in restriction buffer or TE. Solution 1 1 M Sorbitol 20 mM EDTA, pH 8 14 mM b-mercaptoethanol (•-ME) Solution 2 1 M Sorbitol 20 mM EDTA, pH 8 14 mM b-ME 10 mM Tris/HCl pH 7.5 2 mg/ml Zymolase-20T (ICN) (make up fresh) Enzyme solution 6 mg/ml zymolase-20T (ICN) in Solution 1 100% NDS 350 ml H2O, 93 g EDTA 0.6 g Tris base. 1. Add NaOH pellets until pH > 8.0. 2. Dissolve 5 g N-Lauryl-Sarcosine in 50 ml H2O and add to above. 3. PH to 9.5 with NaOH, adjust volume to 500 ml. 4. Store at RT. 2001 GENETICS CORE FACILITIES CATALOG 36 Agarose Solution 1 M Sorbitol University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 45 20 mM EDTA, pH 8 2% Sea Plaque GTG Agarose (FMC) Once melted, cool to 40 C and add •-ME to 14 mM LDS 100 mM EDTA pH 8.0 10 mM Tris/Cl pH 8 1% Lithiumdodecyl sulfate Preparation of ES Cells (Procedure to prepare ES cells for blastocyst injection on Friday) Courtesy of Joanne Thorvaldsen from the Laboratory of Dr. Marisa Bartolomei, Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine. On Monday 1. Seed ES cells onto one well of 24-well plate 1a. Plate mouse embryonic feeder cells (MEFs) onto multiple wells of 24-well gelatinized plate. 1b. After MEFs have attached to plate (2+ hrs), thaw ES cells and seed into one well of the prepared 24-well plate. On Wednesday 2. Split cells as necessary, onto wells of the prepared 24well plate. 2a. If well is confluent (50-60%), split to various densities into three wells of the 24-well plate e.g., approximately 1:2, 3:10 and 1:5 into each well. (See Note below.) The trypsinization method described below works well to ensure single-cell suspension. 2b. If colonies are few or are not growing well, plate trypsinized cells onto one well. In addition, consider plating out an additional vial of ES cells onto another well, to ensure some population of ES cells will be ready for Friday. On Thursday or Friday morning 3. Confirm with Jean Richa that he is prepared for your ES cells (Laboratory Telephone: 215/573-3023, Office Telephone: 215/898-6064). On Friday Note _ The harvested cells need to be ready for injection by noon. _ Two separate wells of ES cells which vary in confluence may be harvested simultaneously to provide Dr. Richa with the most optimum cell density required for the blastocyst injections. 4. Feed cells around 8:00am. 5. Trypsinize cells at around 10:20am. 5a. Wash twice with PBS 5b. Add trypsin (0.25% Trypsin, 1 mM EDTA; 0.2-0.3 ml/ well of 24well dish) and incubate for 5 minutes at 37oC. 5c. To disaggregate the cells, use a plugged Pasteur pipette and pipette cells up and down 5X. Then add 1 ml ES media. With a plugged Pasteur pipette inserted into a P-200 type narrow bore yellow tip, pipette entire cell suspension up and down (15-20X). Note _ It is important to generate a single cell suspension for blastocyst injections. Avoid foaming of the medium! 6. Adsorb MEFs on gelatinized 6-well plate and collect ES cells. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 46 6a. Transfer ES cell suspension to one well of gelatinized 6-well plate which contains 1-2 ml ES medium (final volume 2-3 ml) . 6b. Leave plate in incubator for 1 hour. (The MEFs should sit on the plate after one hour while the stem cells are still in suspension.) 6c. Carefully transfer supernatant to 15 ml conical tube and centrifuge for 5 minutes at 900 rpm (e.g. IEC HNSII Centrifuge). 6d. Discard medium and resuspend the cells in appr. 0.5 ml ES cell medium. To a separate tube, add 5 ml ES cell medium. 7. Deliver 15 ml tube(s) containing ES cells and tube with 5 ml ES cell medium to Jean Richa, Room 202A CRB. (Tubes may be delivered on ice in a Styrofoam box.) Basics for culturing ES cells: _ Colonies should be compact with a smooth outline (see ‘Manipulating the Mouse Embryo’, CSHL Press, Ed. Brigid Hogan et al.(1994)). _ Culture in the presence of LIF (1000U/ml, Gibco/BRL) on mitotically inactivated mouse embryonic feeder (MEF) cells that were plated onto gelatinized plates. Use ES medium as recommended for the specific ES cell-line in use. _ Feed cells daily (and 2-3 hr before cells are trypsinized). _ Do not grow cells for more than 3 days (2 days preferred) without splitting (1:3 to 1:6). Confluent/fast growing cells will need to be split one day after seeding/splitting. _ To prepare gelatinized plates (or wells), cover plate with 0.1% gelatin solution and leave plate in biosafety cabinet for 2 hrs. Remove gelatin and allow gelatin to dry for approximately 10 minutes. (Plates must be completely dry before use or storage at 4o C.) On Animal Husbandry Since all animal maintenance and subsequent breeding of transgenic mouse lines will be carried by the individual investigator, a knowledge of basic mouse husbandry is essential. Here are some basic guidelines and suggestions. _ Mice are born 19-20 days after vaginal plug detection (i.e., successful mating). _ If neonates do not show white stomach (milk) 24 hours after birth, suspect a bad mother; foster mother (lactating female) should be used or pups will die. _ Litters should be weaned at 3 weeks after birth. _ Sexual maturity of females: approximately 6 weeks; that of males; approximately 8 weeks. Maturing males should be housed individually as they tend to become aggressive with one another. _ Watch for water bottle leaks; neglected wet cages can result in the death of animals. _ For animal health problems in your colony or more specialized information, please contact the attending veterinarian of the University Laboratory Animal Resources. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 47 CELL CENTER STOCKROOM Maurice Fluellen, Operations Manager 215/898-2069 fluellen@mail.med.upenn.edu Ian Blake, Clerk Allen Shinn, Clerk Sean-Paul Fenton, Clerk Adam Mancini, Clerk Phineas Walsh, Clerk Val Riordan, Clerk 215/898-2069 Stockroom Hours: Monday - Friday, 9:00 a.m. - 5:00 p.m. Closed 1:00 p.m. - 2:15 p.m. every day Location: D1 Richards Building Mailing Address: 502 Clinical Research Building 415 Curie Boulevard Philadelphia, PA 19104-6145 Telephone: 215/898-2069 Telephone Ordering System: 215/898-5306 Fax: 215/573-3263 E-mail (General Information): cellctr@mail.med.upenn.edu E-mail (Oligo Orders): ccoligo@mail.med.upenn.edu Free special ordering service - pay list price or less • Molecular biology and tissue culture products at discount prices • Free delivery to your lab • Immediate pickup availability • Over 1400 items in stock • Products from 35 different vendors • Special orders of non-stocked items available from 20 vendors • Most special orders available next day • No shipping fees or handling fees, ever • Oligo Nucleotides available, see page 35 • 24 hour ordering via phone system or CellNet U N I V E R S I T Y O F P E N N S Y LVA N I A D E PA R T M E N T O F G E N E T I C S Cell Center Stockroom Opening an Account 1. An account must be established for each Principal Investigator before products or services may be ordered. Once all the required information (see below) has been submitted to the Stockroom, the account will be available for use within one business day. 2. Opening and maintaining an account requires no fees or dues. Purchases are billed on a monthly basis. 3. Three different forms are required to set up a new account. a single Lab Account Information Form, one or more Charge Account Information Forms, and several Lab Member Information Forms. These forms can be found beginning on page xii in the Appendix. After these forms are submitted, each Lab Member will receive an e-mail containing his or her User ID and a randomly generated password to access Stockroom University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 48 ordering systems. Forms may be submitted in person or by fax. Lab Account Information Form — this contains basic contact information for each lab, as well as the Business Administrator’s contact information for billing. Only one copy of this form should be submitted for each principal investigator or lab. Charge Account Information Form — this contains basic FinMIS and grant information. A separate copy of this form should be submitted for every funding source that you wish to use for Stockroom purchases. You must specify a short nickname (two to four words) for the Charge Account, to allow all of us to reference it easily. You may limit access to each Charge Account to only certain Lab Members. You may also specify a Charge Account Phone Code for each Charge Account, to allow you to use any or all of your funding sources with our phone ordering system. Lab Member Information Form — a separate copy of this form should be submitted for each Lab Member who will be placing orders, this includes special orders and oligonucleotide synthesis. It allows you to specify which Charge Accounts should be used by each Lab Member, as well as a unique user code for our phone system. The e-mail address and phone number are required, and each Lab Member receives his or her own unique User ID and password. 4. To change account information: It is very important that account information be kept current. Please use the Lab Account Information Form (page xiii) to update delivery locations and other basic laboratory contact information. To add a new grant or other funding source to your Lab Account, you only need to submit a single Charge Account Information Form, where you may specify which Lab Members should have access to the new funds. New Charge Accounts will be available for purchases within one business day of submission of the form. To obtain access for a new Lab Member within your lab, you only need to submit a single Lab Member Information Form, where you may specify which Charge Accounts the new lab member should use. New Lab Members will be given access within one business day of submission of the form. It is essential that the Business Administrator inform the Stockroom when a Charge Account has expired or when a Lab Member has left the lab. This will avoid unauthorized use of the account. Charge Accounts and Lab Members can be de-activated by the Principal Investigator or Business Adminstrator via fax or e-mail. All de-activations are effective immediately or as soon as possible. The Stockroom cannot be held responsible for unauthorized use of your account if you have not kept your account information current. It is your responsibility to ensure that departed Lab Members are removed from your Lab Account and to keep all passwords secure. The Stockroom will not be responsible for any delays due to incomplete forms. How to Place an Order 1. You can place your order at the Cell Center between University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 49 the hours of 9:00 a.m. and 5:00 p.m., Monday-Friday. Please note the Cell Center is closed daily from 1:00 p.m. to 2:15 p.m.. 2. Automated telephone ordering (898-5306): This system allows order entry through a touch-tone telephone and is available 24 hours a day. NOTE: Please listen for the machine’s acknowledgment that the item number was entered before continuing with your order. Also, if you have a multiline Eagle or Extrom digital phone, press *# before starting to enter the codes to simulate a touch-tone phone. The telephone ordering system uses numeric codes to identify specific Charge Accounts, Charge Account passwords, Lab Members and products. You are free to choose codes that would be easy to remember, but the first digit of a number code cannot be “0”. The Charge Account code and password code can range from 4-9 digits, but Lab Member codes must be 4 digits in length. NOTE: Orders placed through the telephone ordering system are not immediately available to be filled by a staff member. If you have an emergency order, please come to the Stockroom, and a staff member will fill your order except between 1:00 and 2:15 p.m. 3. Only Special Orders may be faxed. All other orders should be placed either by the Stockroom kiosks, CellNet, or via the phone ordering system. Special orders should be faxed using the form located the Appendix, or entered at the Stockroom. Special orders are those items which are not stocked at the Stockroom. Any item that is not stocked at the Stockroom may be special ordered. The following vendors allow special orders and most items are available the next day. Specific information about the availability of special orders can be found in each vendor’s section of the catalog. The Stockroom cannot be responsible for vendor backorders, shipping delays or other circumstances beyond our control. SPECIAL ORDER VENDORS Amersham Biosciences Applied Biosystems Roche Molecular Biochemicals BIO-RAD Clontech Denville Scientific Products / Q-Biogene / TKR Invitrogen (Life Technologies) Perkin Elmer Life Sciences New England Biolabs QIAGEN Roche Molecular Biosystems Sigma Stratagene If there are items or vendors that you frequently use that are not currently stocked by the Stockroom, please notify us. We will be happy to consider such additions. All prices and information in this catalog are subject to change. To Pick Up Your Order Customers may specify each order to be delivered or for pickup at the Stockroom. Orders placed at the Stockroom University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 50 may be picked up immediately. For orders placed remotely, orders submitted before 1:00 p.m. may be picked up at any time after 2:15 p.m. Other orders may be picked up at any time starting at 9:00 a.m. the next day. Please remember that the Stockroom is closed between 1:00 p.m. and 2:15 p.m. every day. Please double-check your order before leaving the Stockroom, or before you sign for your delivery, to make sure that it is complete and correct. Keep the receipt for your records. Occasionally, items on your receipt may be marked “o/s” (temporarily out of stock) or “b/o” (backordered by the vendor). The Stockroom has no control over this situation. It is our policy to notify the user by telephone in this situation and keep a copy of the receipt. As soon as the product becomes available to the Stockroom, we will either deliver it or notify the customer. We cannot exchange or refund opened packages. Delivery Morning deliveries (10:00 a.m. to 1:00 p.m.) include all orders placed before 8:00 a.m. for the following buildings: Route A Route B Chemistry†‡ Abramson CRB BRBI Dulles BRBII/III Founders Kaplan Gates Mudd John Morgan New Bolton Center†‡ Johnson Pavilion Old Vet† Maloney Rosenthal† Penn Tower HUP† Ravdin Wistar† Silverstein Stemmler Towne†‡ Afternoon deliveries (after 2:30 p.m.) include all orders placed before 1:00 p.m. for the following buildings: Route A Route B Route C CRB Abramson Anatomy/Chemistry John Morgan BRB Goddard Johnson Pavilion BRB II/III Richards Stemmler Wistar† Leidy Levy† VHUP† There are never any charges for delivery, shipping or handling for any Stockroom order. Labs are limited to ten (10) bottles of media per day. Special arrangements may be made with the Stockroom for larger orders. Please call 898-2069 for more information. † deliveries to these buildings are limited to 5 bottles of media per lab per day and may be limited or suspended during inclement weather at the Stockroom’s discretion. ‡ Tuesday/Thursday morning only or by special request. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 51 OLIGONUCLEOTIDE SYNTHESIS Invitrogen (Life Technologies) Technical support: 800/828-6686 Internet: http://www.lifetech.com Pricing • 10 nmole scale (>2 OD) - $0.25 per base • 50 nmole scale (>2 OD) - $0.41 per base • 200 nmole scale (>10 OD) - $0.90 per base • 1000 nmole scale (>50 OD) - $1.95 per base • Setup fee: No charge (Desalted). • Setup charges: Cartridge 50/200nm $24.00 Cartridge 1000nm $30.00 HPLC 200nm $52.75 HPLC 1000nm $90.00 PAGE 200nm $65.00 PAGE 1000nm $75.00 • No delivery or shipping fees • Delivery: = 45 bases - 2nd business day; > 60 bases, 5’ Phosphorylation, Iosine, Uracil or Cartridge Purity - 3rd business day > 60 bases, 1000 nmole scale, 5’ modifications (except Phosphorylation) or 5’ Oligos - within 5 business days HPLC and PAGE purity - within 6 business days • Cell Center must receive order by 2:30p.m. (fax or email) • Modifications: 5’Phosphorylation: 50/200nm - $25 1000 nm - $56 5’Biotin: 50/200 nm - $49 1000 nm - $110 Inosine: 10/50/200 nm - $8 1000 nm - $16 Mixed bases: N/C Uracil: 10/50/200 nm - $7 1000 nm - $10 Technical Information LTI Custom Primers are synthesized on a highly automated computer controlled parallel array synthesizer using standard phosphoramidite chemistry. Each order is double entered into the system ensuring correct input. Coupling efficiencies are monitored throughout synthesis on each oligonucleotide by trityl analysis. Five levels of purity are currently available: deprotected only; deprotected/desalted by C18 spin-column chromatography; cartridge; HPLC and PAGE. A comprehensive certificate of analysis accompanies each oligonucleotide indicating quantity, molecular weight, extinction coefficient, sequence, and melting temperature. Ordering Information Place your order by: FAX: use the appropriate forms located in the Appendix and fax to 215/573-3263 by 2:30 p.m. E-Mail: use the following format and send to ccoligo@mail.med.upenn.edu by 2:30 p.m.. Invitrogen Procedure for Sending Custom Oligo Orders Via E-mail _ Please follow instruction exactly (no additional or University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 52 fewer spaces). _ “Subject” line must indicate LTI OLIGO. LTI User Name ID Password (non numerical) Charge Account Phone Number To Be Delivered? (Y or N) Date * Researcher Name Primer Name Sequence (All capitals please, limit 100 bases 5’ to 3’) 50, 200, or 1000 (nmole scale) P, G, R, L, B, H, T, F or # P if 5’ Phosphorylation G if 5’ Fluorescein R if 5’ Rhodamine L if 5’ Primary Amine B if 5’ Biotin H if 5’ HEX T if 5’ TET F if 5’ FAM # = none D, S, O, P or H D = Deprotected S = de(S)alted O = Cartridge P = PAGE, H= HPLC * Next Primer (same format as above) * Last Primer (same format as above) Sigma-Genosys Technical support: 800/234-3693 Pricing • 3OD Select - $0.30 per base 50 nmole scale - $0.70 per base 200 nmole scale - $1.30 per base 1000 nmole scale - $2.20 per base • Setup fee: No charge (Desalted). • Setup charges: Cartridge 50/200nm $20.00 Cartridge 1000nm $32.20 HPLC 50/200nm $60.00 HPLC 1000nm $88.55 PAGE 50/200nm $60.00 PAGE 1000nm $88.55 • No delivery or shipping fees • Delivery: = 35 bases - 2nd business day; > 35 bases - 3rd business day; 5’ modifications or 5’ Oligos - within 5 business days HPLC and PAGE purity - within 5 business days • Cell Center must receive order by 2:30p.m. (fax or email) • Modifications: 5’Fluorescein: 50/200nm - $40.25 1000 nm - $80.50 5’Phosphorylation: 50/200nm - $20 1000 nm - $40.25 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 53 5’Biotin: 50/200 nm - $40.25 1000 nm - $80.50 Inosine: 50/200 nm - $6.45 1000 nm - $12.00 Mixed bases: N/C Please call the Stockroom for more information regarding Sigma-Genosys oligos. Technical Information Sigma-Genosys oligos are synthesized using our patented AbacusTM platform which yields a higher coupling efficiency than most competitor platforms. We offer 4 levels of purification: desalt, reverse phase cartridge (RP1), PAGE and reverse phase HPLC. Each and every oligo is physically tested by gel electrophoresis and spectrophotometer analysis. Additionally, each synthesis run is statistically tested using MALDI-ToF mass spectrometry analysis. At Sigma-Genosys, quality is not just a goal - it is a guarantee. In addition to our stringent quality analysis, you will receive a Quality Assurance Certificate that includes quantitated yield, melting temperature, MW and µg/OD with every oligo. You will also receive an additional label that can be used in your records or on an additional vial. All this at no extra charge. Ordering Information Place your order by: FAX: use the appropriate forms located in the Appendix and fax to 215/573-3263 by 2:30 p.m. E-Mail: use the following format and send to ccoligo@mail.med.upenn.edu by 2:30 p.m. Sigma-Genosys Procedure for Sending Custom Oligo Orders Via E-mail _ Please follow instruction exactly (no additional or fewer spaces). _ “Subject” line must indicate GENOSYS OLIGO. SIGMA GENOSYS User Name ID Password (non numerical) Charge Account Phone Number To Be Delivered? (Y or N) Date * Researcher Name Oligo Name Sequence (All capitals please, limit 110 bases 5’ to 3’; 3OD 10-35 bases) 3 OD Select, 50, 200, or 1000 (nmole scale) DST, RP1, PAGE or HPLC DST = Desalted RP1 = Cartridge PAGE HPLC NOTES (Define any special modifications) * Next Primer (same format as above) * Last Primer (same format as above) University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 54 PRIMERS & PROBES Applied Biosystems Technical Support: 800/345-5224 Internet: http://www.pebiosystems.com Product Offering & Pricing _ Flat Rate plus $3.50 Setup fee _ No Delivery or Shipping Fees _ Cell Center must receive order by 12:30p.m. _ Desalted & Deprotected _ 40nm & 200nm scale Prism Primers & Prism Primer Pairs delivered in 4 days. Automatic purification included. Labels available include: 6-FAM, HEX, NED, or TET _ 40nm & 200nm TaqMan Probes and Sequence Detection Primers delivered together in 5 days. HPLC purification included. Labels available include: 6FAM,VIC, or TET on 5’ end; TAMARA on 3’ end. ABI Prism® Primers 450007 40nm 5’-Fluorescent-labeled oligo. Maximum of 30 bases. 4000pmol purified. Flat Rate: $78.00 450006 200nm 5’-Fluorescent-labeled oligo.Maximum of 30 bases. 40,000 pmol purified. Flat Rate: $125.00 450017 1um 5’-Fluorescent-labeled oligo.Maximum of 30 bases. 300,000pmol. Flat Rate: $158.00 Purified yeild of 150,000pmol Extra 2 days HPLC Charge: $86.00 ABI Prism® Primer Pairs 450056 40nm Primer Pair (one 5’Fluorescent-labeled & one unlabeled up to 30mers.) 4000pmol each primer purified. Flat Rate: $100.00 450096 40nm Individual Loci for ABI Linkage Mapping Set 5cM Map. 4000pmol of each primer purified. Flat Rate: $52.50 450059 200nm Primer Pair (one 5’Fluorescent-labeled & one unlabeled up to 30mers.) 40,000pmol each primer purified. Flat Rate: $157.00 450062 1um Primer Pair (one 5’Fluorescent-labeled & one unlabeled up to 30mers.) 300,000pmol each primer. Flat Rate: $234.00 Extra 2 days HPLC Charge: $86.00 Tailed Primer Pairs: _Adding a 7 base sequence[primer tailing] on the reverse primer. Same specifications per scale as above [F] primer pairs. 4304979 40nm scale tailed Primer Pair Flat Rate: $103.20 4304981 200nm scale tailed Primer Pair Flat Rate: $159.10 4304982 1um scale tailed Primer Pair University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 55 Flat Rate: $236.50 TaqMan® Probes & Sequence Detection Primers 450025 40nm TaqMan Probe, 5000-6000pmol Flat Rate: $290.00 4304970 40nm Sequence Detection Primer, 4000pmol Flat Rate: $25/primer 450024 200nm TaqMan Probe, 15000-25000pmol Flat Rate: $540.00 4304971 200nm Sequence Detection Primer, 80000pmol Flat Rate: $35/primer 450003 1um TaqMan Probe, 50,000-100,000pmol Flat Rate: $750.00 450005 1um Sequence Detection Primer, 300,000pmol Flat Rate: $62/primer Ordering Information _Place your order by e-mail using the following format and send by 2:30p.m. to ccoligo@mail.med.upenn.edu Subject Line must indicate ABI OLIGO. ABI User Name ID Password (non numerical) Charge Account Phone Number To Be Delivered? (Y or N) Date * Example: Part# 450007 Sequence name oligo 1 Sequence NEDagtccgcgtcgatctcga For labels please use the following nomenclature: NED,HEX, FAM, TET, VIC, Biotin, TAMARA For redundancies, use the following IUB Group Codes: M=(AC); R=(AG); W=(AT); S=(CG); Y=(CT); K=(GT); V=(AGC); H=(ACT); D=(AGT); B=(CGT); N=(AGTC). Stocked Products - Product Class Listings .................................................. Chemicals and Reagents • (CH).......................................................................................... Competent Cells • (CE) ...................................................................................................... Film and Membranes • (FM) .............................................................................................. Immunological Reagents • (IM) ......................................................................................... Miscellaneous Products • (MS) .......................................................................................... Molecular Biology Kits • (KT) ........................................................................................... Molecular Biology Products • (MB) .................................................................................. Non-radioactive Kits and Reagents • (NR). ....................................................................... Other Enzymes • (OE) ........................................................................................................ PCR Reagents and Nucleotides • (PC) ............................................................................... Precast Gels • (PG) ............................................................................................................ Protein & Molecular Weight Markers • (MR) .................................................................... Purification Kits and Columns • (CO)................................................................................ Restriction Enzymes • (RE) ............................................................................................... Sera • (SM) ........................................................................................................................ Tissue Culture Media and Reagents • (TC) ....................................................................... Stocked Products - Listed by Vendor ........................................................... Amersham Biosciences • (AMR) .................…………….................................................. Applied Biosystems • (PEC) ............................................................................................ 57 58 57 63 64 68 65 66 69 69 71 74 67 62 74 78 78 80 80 101 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 56 Bio-Rad Laboratories • (RAD) ........................................................................................ Biowhittaker Molecular Applications • (BMA) ................................................................ Cambrex Biosciences • (FMC) ......................................................................................... Cell Center Services • (CCS) ............................................................................................. Clontech Laboratories • (CLN) .......................................................................................... Collaborative Biomedical Products • (COL) ..................................................................... Denville Scientific • (DEN) ............................................................................................... Difco Laboratories • (DIF) ................................................................................................. Enzo Life Sceinces • (ENZ) ............................................................................................... Epicentre Technologies • (EPI) ......................................................................................... Fisher Scientific • (FSH) .................................................................................................... Fuji Film • (FUJ) ................................................................................................................ Gemini Bioproducts • (GEM) ............................................................................................ Hyclone Laboratories • (HYC) .......................................................................................... IGEN International (Fisher Scientific) • (IGN) ................................................................. Invitrogen (Life Techologies) • (INV) ............................................................................... Kodak • (KOD) .....................................................................................…......................... Marligen Biosciences • (MLG)........................................................................................... Mediatech (Fisher Scientific) • (MED) .............................................................................. Micron Separations Inc. (Fisher Scientific) • (MSI) .......................................................... Millipore • (MIL) ................................................................................................................ National Diagnostics • (ND) ............................................................................................... New England Biolabs • (NEB) ........................................................................................... Perkin Elmer (New England Nuclear) • (NEN) ................................................................ Polaroid Film • (POL) ....................................................................................................... Promega • (PRO)....................................................................................…........................ Q-Biogene • (DEN) ............................................................................................................ Qiagen • (QIA) .................................................................................................................. Rainin Instruments • (RAI) ............................................................................................... Roche Diagnostics • (BMB) .........................………………………………………......... Sigma-Aldrich • (SIG) ...................................................................................................... Stratagene • (STR) ..................................................................................…...................... TKR Bioproducts • (DEN) ................................................................................................ USB Corporation • (USB) ............................................................................................... Index of Stocked Products .......................................................................... 106 85 86 86 87 87 88 89 89 89 89 91 91 91 91 91 97 97 97 98 98 99 99 101 102 103 88 105 107 82 107 108 88 109 109 STOCKED PRODUCTS U N I V E R S I T Y O F P E N N S Y LVA N I A D E PA R T M E N T O F G E N E T I C S COMPETENT CELLS .............................................................................................................. CE Mfgr. Mfgr No STR 230245 STR 230255 STR 230240 STR 230250 LTI 18263-012 LTI 18265-017 LTI 18290-015 Code 5428 5430 5427 5429 4118 4119 5423 Item Description BL21-Codon Plus (DE3)-RIL Competent Cells BL21-Codon Plus (DE3)-RP Competent Cells BL21-Codon Plus-RIL Competent Cells BL21-Codon Plus-RP Competent Cells DH5alpha™ Library Efficiency Comp Cells DH5alpha™ Subcloning Efficiency Comp Cells ElectroMAX DH10B Efficiency Comp Cells Unit ea ea ea ea 1 ml 2 ml 5x.1ml CC Price $ 196.77 $ 196.77 $ 196.77 $ 196.77 $ 94.88 $ 43.69 $ 156.66 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 57 LTI PRO PRO LTI LTI LTI INV LTI INV INV INV STR STR STR STR STR STR STR 11319-019 L2011 L2001 12034-013 18258-012 10268-019 C862003 12297-016 C2020-03 C4040-03 C3030-03 200152 200238 200315 200228 200314 200236 200150 5426 4246 4247 5458 5425 5474 4637 4622 4334 5445 4335 4550 4129 5418 4258 4548 4130 5517 ElectroMAX DH5alpha-E Efficiency Comp Cells HB101 High Efficiency Competent Cells JM109 High Efficiency Competent Cells Max Efficiency DH5@T1 Competent Cells MAX Efficiency DH5alpha Efficiency Comp Cells Max Efficiency STBL2 One Shot MACH 1 Comp eColi (LTI Shelf) One Shot® MAX Efficiency® DH5a®-T1® OneShot™ INVaF' Competent Cells OneShot™ TOP10 Competent Cells OneShot™ TOP10F' Competent Cells SURE® 2 Supercompetent Cells SURE® Competent Cells XL 10-Gold Ultracompetent Cells XL-1 Blue Electroporation Competent Cells XL10-Gold Ultracompetent Cells XL-1-Blue Supercompetent Cells XL-2 Blue Ultra Competent Cells 5x.1ml 1 ml 1 ml 1 ml (5 x 0.2ml) 5x.1ml 1 ML 20 rxn 20 x 50ul 21x50µl 21x50µl 21x50µl ea 5x0.2ml 10 x 100µl 5x80µl 5x100ul 5x0.2ml ea $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 154.63 58.83 67.61 123.50 157.71 148.43 235.31 243.98 243.61 238.08 235.31 156.21 152.30 194.93 161.25 129.39 146.57 183.82 CHEMICALS AND REAGENTS .............................................................................................. CH Mfgr. Mfgr No Code Item Description MED MT46-010-CM 1290 10X TAE MED MT46011CM 1291 10X TBE DEN CS-5585-28 3306 20% SDS Solution MED MT46020CM 1292 20X SSC Buffer LTI 10812-014 5453 2nd Strand Buffer RAD 1610154 2392 30% Acrylamide/Bis 19:1 RAD 1610156 2393 30% Acrylamide/Bis 29:1 RAD 1610158 2394 30% Acrylamide/Bis 37.5:1 SIG A8456-100mg 5913 4-(2-Aminoethyl)Benzenesulfonyl*Fluoride Hydrochloride RAD 1610144 2389 40% Acrylamide/Bis 19:1 RAD 1610146 2390 40% Acrylamide/Bis 29:1 RAD 1610148 2391 40% Acrylamide/Bis 37.5:1 NDS EC850 2101 AccuGel™ 19:1 AMR 27-6200-01 2340 Acetyl CoA BMB 101893 2102 Acetyl CoA BMB 1685791 2104 Acrylamide FSH BP170500 2224 Acrylamide RAD 1610101 2347 Acrylamide LTI 30391-023 4185 Agar - Bacterial Select DIF DF0145170 5889 Agar - Granulated FSH BP1423500 4308 Agar - Granulated LTI 15510-019 2105 Agarose LTI 15510-027 2106 Agarose FMC 50100 2234 Agarose (LMP) SeaPlaque® FMC 50101 2263 Agarose (LMP) SeaPlaque® LTI 10975-035 1289 Agarose 1000 FMC 50090 2321 Agarose NuSieve® 3:1 FMC 50080 2233 Agarose NuSieve® GTG® FMC 50004 2236 Agarose SeaKem® LE Unit CC Price 1L $ 1L $ 500ML $ 1L $ 0.5 ml $ 500 ml $ 500 ml $ 500 ml $ 100 mg $ 500 ml $ 500 ml $ 500 ml $ 450 ml $ 10 mg $ 10 mg $ 500 g $ 500 g $ 500 g $ 500 g $ 500 g $ 500 g $ 100 g $ 500 g $ 125 g $ 25 g $ 100G $ 125 g $ 125 g $ 500 g $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 58 12.70 11.65 30.10 9.51 53.89 39.32 39.32 39.32 66.71 45.57 45.57 45.57 43.76 53.56 52.56 95.70 74.59 99.19 64.27 59.09 68.93 73.39 256.80 289.24 105.93 181.90 223.98 278.01 285.54 FMC FSH RAD CCS SIG BMB FSH USB BMB RAD FSH RAD BMB INV RAD FSH SIG RAD DIF FSH BMB BMB FSH CCS FSH BMB BMB RAD USB AMR FSH SIG BMB FSH RAD FSH SIG CLN CLN CLN CLN SIG LTI FSH LTI SIG SIG AMR AMR BMB FSH AMR LTI BMB FSH 50000 BP179100 1610700 2111 A6279-10ml 1383221 BP1610100 US12387 874515 1610786 BP17125 1610201 1685830 R21001 1610710 BP168500 B2518-100mg 1610404 DF0230173 BP1424500 757306 757292 BP210500 2117 BP339500 1873580 1836153 1610400 70468 17-0340-01 BP1585100 D2650-5x5ml 100032 BP1725 1610611 BP2311 D2650-100ml 630415 630414 630413 630416 D9891-25g 15575-020 BP120500 15585-011 E1510 E4391 17-1440-03 17-1440-02 1814320 BP227500 17-0756-01 15514-011 100149 BP3811 2235 4337 2352 2111 5912 2379 2405 2113 2303 6058 4338 2350 2114 4635 5908 2406 2446 5911 5890 4309 2115 2116 2226 2117 4339 5522 5521 5909 2121 2212 4440 5914 2122 2239 6060 2407 6062 5992 5989 5981 5991 2445 2283 2408 2277 2375 2376 2126 2125 2127 2227 2213 2130 2131 2230 Agarose SeaKem® LE 125 g Ammonium Persulfate 100 g Ammonium Persulfate 10 g Ampicillin - 25mg/ml 10 ml Aprotinin from Bovine Lung 10 ml BCIP 3ml/150mg BCIP 100 mg BCIP 100 mg Bestatin 10 mg Bio-Safe Coomasie Stain 1L Bis-Acrylamide 25 g Bis-Acrylamide 50 g Bis-Acrylamide 25 g Blasticidin 50 MG BME (2-mercaptoethanol) 25ml Boric Acid 500 g BOVINE SERUM ALBUMIN MOLECULAR BIOLOGY GRADE 100g Bromophenol Blue 10g Casamino Acids 500 g Casamino Acids 500 g Cesium Chloride 1 kg Cesium Chloride 100 g Cesium Chloride 500 g Chloramphenicol 34 mg/ml 10 ml Citric Acid 500 g Complete EDTA Free 20 Tabs EA Complete Mini 25 Tabs EA Coomassie™ Brilliant Blue R-250 10g Denhardt's Solution 50 ml Dextran Sulfate 100 g Dextran Sulfate 100 g Dimethyl Sulfoxide Hybri-Max® Sterile*Filtered 25 ml Dithiothreitol (DTT) 5g Dithiothreitol (DTT) 5g Dithiothreitol (DTT) 5g DMSO 1L DMSO 100 ml DO Supplement-His 1 ea DO Supplement-Leu 1 ea DO Supplement-Trp 1 ea DO Supplement-Ura 1 ea DOXYCYCLINE HYDROCHLORIDE 25g EDTA 0.5M (4 x 100ml) 400 ml EDTA Disodium Salt Dihydrate 500 g Ethidium Bromide (ETBR) 10mg/ml 10 ml Ethidium Bromide Aqueous Solution 10mg/ml 10 ml Ethidium Bromide Tablets 10mg 10 ea Ficoll-Paque® Plus solution 6x500ml Ficoll-Paque® solution 6x100ml Formamide 500 ml Formamide 500 ml Glutathione Sepharose 10 ml Glycerol 500 ml Glycine 1 kg Glycine 1 kg $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 59 111.25 15.24 10.72 10.00 80.37 37.65 18.65 24.34 99.62 51.41 19.44 28.60 26.67 140.80 16.98 17.59 95.16 27.70 51.84 46.88 261.21 49.42 131.63 8.00 15.72 225.13 96.48 16.08 37.00 119.29 87.37 29.82 54.91 42.03 52.48 30.36 84.34 34.29 37.07 34.29 34.29 174.74 23.91 30.64 28.47 34.25 31.02 337.40 99.23 36.08 30.90 159.09 18.18 40.79 29.35 LTI RAD LTI FSH BMB FSH LTI INV INV INV BMB FSH FSH CCS CCS CCS LTI SIG DIF FSH CCS MED SIG DIF FSH DIF FSH BIO BMB SIG FSH CLN CLN BMB USB INV INV INV INV INV INV INV INV RAD BMB BMB DIF FSH AMR SIG SIG FSH LTI BMB BMB 15527-013 1610718 15502-016 BP221250 1685929 BP310100 15630-080 Q602-20 Q612-20 Q621-20 724815 BP16201 BP162010 6005 6006 6007 22700-025 L7533 DF0445174 BP1425500 6004 MT46-050-CM L7658-1kg DF0402170 BP1427500 DF0446173 BP1426500 3002-031 1017128 L2884-10mg BP214500 630412 630411 1383213 19535 NP00061 NP0005 NP0007 NP0002 NP0001 NP0004 NP0006 LA0041 1610202 1429868 1359053 DF0118170 BP1420500 17-0891-01 H7033-125ml H7033-1L BP226500 15513-039 100995 1814303 2132 2354 2133 4442 2134 4441 4445 6092 6094 6095 2145 2238 5299 6005 6006 6007 4186 5916 5891 4310 6004 1294 5917 5893 4311 5892 4312 2420 2146 6063 2410 5905 5903 2378 2156 4011 6081 6082 6083 6084 6085 6086 6087 2395 2377 2172 5886 4313 2173 6066 5448 2231 2176 2237 2174 Glycine Glycine Guanidine HCL Guanidine Isothiocyanate Guanidine Thiocyanate HEPES HEPES Buffer Solution (1M) imMedia™ Amp Blue imMedia™ Kan Blue imMedia™ Zeo Agar IPTG IPTG IPTG LB + Ampicillin (50ug/ml) Agar Plates LB + Kanamycin (50ug/ml) Agar Plates LB + Tetracycline (20ug/ml) Agar Plates LB Agar LB Agar - EZMix® LB Agar - Miller LB Agar - Miller LB Agar Plates LB BROTH LB Broth - EZMix® LB Broth - Lennox LB Broth - Lennox LB Broth - Miller LB Broth - Miller LB Medium Capsules Leupeptin Leupeptin Hemisulfate from Microbial Source Magnesium Chloride Minimal SD Agar Base Minimal SD Base NBT (Nitro Blue Tetra) NBT (Nitro Blue Tetra) NP Transfer Buffer (20X) NuPAGE® Antioxidant NuPAGE® LDS Sample Buffer (4X) NuPAGE® MES SDS Running Buffer (20X) NuPAGE® MOPS SDS Running Buffer (20X) NuPAGE® Sample Reducing Agent (10X) NuPAGE® Transfer Buffer (20X) NuPAGE® Tris-Acetate SDS Running Buffer (20X) PDA Pefabloc® SC Pepstatin (was #600160) Peptone Peptone Percoll® Perfect Hyb Plus Hybridization Buffer Perfect Hyb Plus Hybridization Buffer Phenol Phenol Saturated (Buffer) Phenol Saturated (Tris) Phenol (was 100300) 500 g 1 kg 500 g 250 g 500 g 100 g 100 ml ea ea ea 1g 1g 10 g 10 pl 10 pl 10 pl 500 g 6 x 500 ml 500 g 500 g 10 pl 1L 1 kg 500 g 500 g 500 g 500 g 1 kg 25 mg 10 mg 500 g 1 ea 1 ea 3 ml 1g 1L 15ml 10ml 500ml 500ml 250ul 125ml 500ml 10 g 100 mg 10 mg 500 g 500 g 1L 125 ml 1L 500 g 100 ml 100 ml 500 g $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 60 23.53 29.49 65.42 58.61 135.71 32.36 26.77 225.64 177.96 276.96 32.16 27.17 202.45 10.00 10.00 10.00 47.15 21.68 44.71 54.23 10.00 13.78 122.11 20.43 26.88 20.43 26.94 107.64 105.11 57.64 22.37 139.00 92.66 36.87 59.94 100.04 15.28 7.64 43.07 43.07 7.64 13.43 41.22 75.96 90.99 47.07 41.30 46.06 226.82 75.03 427.49 63.92 58.24 48.63 92.56 FSH FSH FSH FSH SIG FSH BMB SIG SIG SIG SIG SIG SIG SIG BMB BMB BMB FSH FSH BMB NDS NDS NDS LTI NDS NDS NDS NDS NDS NDS BMB FSH RAD BMB LTI LTI BMB FSH LTI USB RAD BMB RAD SIG CCS SIG SIG BMB BMB RAD LTI LTI FSH FSH LTI BP1750I100 BP1750I400 BP1752I100 BP1752I400 P7626-5g BP366500 1697498 P8465-5ml P8465-25ml P2714 P8340-1ml P8340-5ml P8849-5ml P8215-1ml 3115879 1373200 1000144 BP1700100 BP1700500 3115828 EC890 EC-892 EC-893 15553-027 EC835 EC830 EC840 EC833 EC836 EC838 1667262 BP166500 1610301 1666681 15557-036 15591-035 100168 BP2201 15558-026 US75824 1610733 100139 1610800 T9179 2197 T9424-25ml T9424-100ml 604205 604203 1610719 15567-027 15568-025 BP1521 BP13381 15506-017 5300 5301 5302 5303 5918 2411 2345 2403 2404 2398 2401 2402 6065 2399 5136 2444 5518 5211 5304 5526 2177 2341 2342 2284 2179 2180 2181 2182 2184 2183 2185 2232 5910 2186 2279 2285 2189 4443 2275 2297 2355 2196 2351 5922 2197 5923 6064 2198 2199 2353 2281 2282 4444 5305 2202 Phenol Saturated pH 6.6 Phenol Saturated pH 6.6 Phenol/Chloroform/Isoamyl Alcohol Phenol/Chloroform/Isoamyl Alcohol Phenylmethylsulfonyl Fluoride Potassium Chloride Protease Inhibitor Cocktail Tablets (Complete) Protease Inhibitor Cocktail-Bacterial Protease Inhibitor Cocktail-Bacterial Protease Inhibitor Cocktail-General Protease Inhibitor Cocktail-Mammalian w/o Metal Chelators Protease Inhibitor Cocktail-Mammalian w/o metal chelators Protease Inhibitor Cocktail-Poly His Tagged Protease Inhibitor Cocktail-Yeast/Fungal Proteinase K Proteinase K Proteinase K Proteinase K Proteinase K Proteinase K Solution (20 mg/ml) ProtoGel® (30% Acrylamide/0.8% Bis) ProtoGel® Buffer ProtoGel® Stacking Buffer SDS 10% (4x100ml) SequaGel® Buffer SequaGel® Concentrate (20% Acryl.) 19:1 SequaGel® Diluent SequaGel® Sequencing System SequaGel®-6 6% Monomer Sequencing Soln (includes EC-841) SequaGel®-8 8% Monomer Sequencing Soln (includes EC-841) Sodium Dodecyl Sulfate Sodium Dodecyl Sulfate (SDS) Sodium Dodecyl Sulfate (SDS) SSC 20X SSC 20X SSPE 20X Sucrose Sucrose TAE 10X Taurine TBE 10X TEMED TEMED Terrific Broth (Modified)-EZMix® Tetracycline 12.5mg/ml Tri Reagent Tri Reagent Tris Tris Tris TRIS pH 7.5 TRIS pH 8.0 Tris Base TRIS EDTA Solution 100X Tris Hydrochloride 100 ml 400 ml 100 ml 400 ml 5g 500 g 20 tabs 5 ml 25 ml 1 btl 1 ml 5 ml 5 ml 1 ml 100 mg 25 ml 500 mg 100 mg 500 mg 5 ml 1L 450 ml 200 ml 400 ml 200 ml 1L 1L 1 kit 1L 1L 100 g 500 g 100g 4L 4L 4L 1 kg 1 kg 4L 1 kg 1L 50 g 5 ml 6 x 500ml 10 ml 25 ml 100 ml 1 kg 500 g 1 kg 1L 1L 1 kg 1L 500 g $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 61 33.30 71.96 30.24 72.45 43.08 15.18 225.13 29.43 118.05 24.85 29.74 118.82 118.05 29.49 59.61 313.77 61.97 61.80 285.55 90.99 72.98 19.36 16.00 25.51 10.00 54.20 20.90 85.58 37.44 39.98 29.81 61.80 25.91 80.80 49.53 79.75 32.16 20.57 58.24 115.06 21.02 32.16 13.40 67.48 8.00 24.11 75.55 57.26 36.08 55.40 17.73 19.28 51.75 55.41 50.10 RAD FSH FSH BMB SIG SIG SIG SIG SIG LTI MED FSH DIF RAD FSH LTI FSH BMB BMB CCS DIF FSH LTI 1610732 BP152500 BP13071 789704 T6066-1Kg T1503-1kg T3253-1kg T2444 T2694 15596018 MT25-900-CI BP1421500 DF0123173 1706531 BP169500 15505-050 BP5611 745740 651745 2207 DF0127179 BP1422500 22712-020 2397 2412 2413 2203 5921 5919 5920 1287 1288 5457 1293 4314 5887 6059 2240 2204 2409 2205 2206 2207 5888 4315 4189 TRIS/Glycine/SDS 10X Buffer Tris-Base Tris-Glycine (10X) Triton® X-100 TRIZMA® Base Molecular Biology Reagent Grade TRIZMA® Base Reagent Grade TRIZMA® Hydrochloride Reagent Grade Trizma-Hcl, 1 M Stock Solution, pH 7.6 Trizma-Hcl, 1 M Stock Solution, pH 8.0 Trizol Trypan Blue Solution Tryptone Tryptone Peptone Tween 20 Urea Urea Water(Sterile Nuclease Free) X-gal X-gal X-gal Yeast Extract Yeast Extract YT Broth 2X 1L 500 g 1L 100 ml 1 kg 1 kg 1 kg 1L 1L 200 ml 100ML 500 g 500 g 100 ml 500 g 2 kg 1L 1g 250 mg 1g 500 g 500 g 500 g $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ PURIFICATION KITS AND COLUMNS .................................................................................. CO Mfgr. Mfgr No QIA 34698 CLN 636055 CLN 636060 QIA 301425 QIA 12362 INV K1593-02 BIO 1102-200 BIO 1101-200 BIO 1101-400 BIO 1001-400 BIO 1001-600 SIG May-00 BMB 1754777 BMB 1754785 QIA 12643 BIO 2055-400 INV K1520-02 AMR 27-5325-01 AMR 27-5330-01 AMR 27-5120-01 AMR 27-5130-01 AMR 17-0855-02 QIA 30410 PRO Z5400 AMR 27-5335-01 QIA 51104 QIA 51304 Code 4603 4533 4536 4601 4502 5221 4614 4450 4569 4269 4448 4539 4512 4513 4602 4451 5220 4467 4507 4509 4299 4148 4503 4528 4297 4318 4319 Item Description Anti-His Antibody Selector Kit (3 x 3 µg) CHROMA SPIN-10 Columns CHROMA SPIN-100 Columns Effectene Transfection Reagent (1 ml) EndoFree Plasmid Maxi Kit (10) FastTrack® 2.0 Kit GeneClean Turbo GeneClean® DNA Purification Spin Kit GeneClean® DNA Purification Spin Kit GeneClean® II DNA Purification Kit GeneClean® III DNA Purification Kit Genelute Agarose Spin Column High Pure Plasmid Isolation Kit High Pure Plasmid Isolation Kit Hispeed Midi Kit (25) 45 minutes/200ug Lambda Quick! Phage DNA Isolation Spin Kit Micro-FastTrack™ Kit MicroSpin G-25 Columns MicroSpin G-50 Columns MicroSpin S-200 HR Columns MicroSpin S-300 HR Columns NICK® Columns Ni-NTA Superflow PolyATtract® System 1000 RNA Isolation System ProbeQuant™ G-50 Micro Columns QIAamp™ Blood Kit QIAamp™ DNA Mini Kit Unit 3x3ug 1 ea 1 ea 1 ml kit 6 rxn 50 prp 50 prp 100 prp 300 prp 600 prp 70 ea 50 col 250 col 45min/200ug 25 prp 20 rxn 50 col 50 col 50 col 50 col 50 col 25 ml 1 kit 50 col 50 prp 50 prp CC Price $ 70.88 $ 112.10 $ 112.12 $ 197.38 $ 211.73 $ 322.10 $ 57.36 $ 78.53 $ 139.44 $ 123.53 $ 180.01 $ 76.00 $ 47.85 $ 206.30 $ 220.71 $ 100.60 $ 369.52 $ 119.71 $ 119.71 $ 119.71 $ 119.71 $ 219.73 $ 221.60 $ 279.25 $ 119.71 $ 90.61 $ 100.48 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 62 19.66 32.76 31.44 25.10 72.53 71.70 113.46 16.69 16.69 149.82 6.63 37.83 36.39 35.81 21.21 67.36 17.59 147.47 86.29 65.00 51.68 54.35 33.58 QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA QIA RAD RAD RAD BMB BMB BMB BMB BMB QIA QIA BIO QIA PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO PRO 30210 20021 12191 12162 12163 12165 12181 12143 12123 27106 27104 28144 28706 28704 28304 28104 79654 7326130 7326120 7326100 1273922 1273990 1273965 1273973 1274015 74106 74104 2070-400 301305 PRA7280 PRA7290 PRA7270 A7300 PRA7640 A7211 PRA7141 A7170 A7100 A7510 A1330 A1460 4316 4158 4564 4159 4317 4565 4281 4161 4163 4566 4203 4567 4568 4204 4504 4255 4282 5437 5438 4522 4165 4166 4167 4326 4168 4595 4256 4214 4547 4142 4143 4144 4145 4195 4292 4293 4147 4146 4291 4529 4531 QIAexpress™ Ni-NTA Agarose Kit QIAEX™ II Gel Extraction Kit QIAGEN Plasmid Giga Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Mega Kit QIAGEN Plasmid Midi Kit QIAGEN Plasmid Mini Kit QIAprep Spin Miniprep Kit QIAprep™ Spin Miniprep Kit QIAquick 8 PCR Purification Kit QIAquick Gel Extraction Kit QIAquick™ Gel Extraction Kit QIAquick™ Nucleotide Removal Kit (50) QIAquick™ PCR Purification Kit QIAshredder™ Quantum Maxi-Prep Kit Quantum Midi-Prep Ket Quantum™ Prep Plasmid Miniprep Quick Spin™ G25 DNA Quick Spin™ G25 RNA Quick Spin™ G50 DNA Quick Spin™ G50 DNA Quick Spin™ G50 RNA RNeasy Mini Kit RNeasy™ Mini RNA Kit RPM™ Rapid Pure Plasmid Miniprep Kit SuperFect Transfection Reagent (1.2 ml) Wizard™ DNA Clean-up System Wizard™ Lambda System Wizard™ Maxipreps System Wizard™ Megapreps System Wizard™ Midipreps System Wizard™ Minicolumns Wizard™ Miniprep DNA Purification Resin Wizard™ PCR Preps System Wizard™ Plus Miniprep System Wizard™ Plus Miniprep System Wizard™ Plus SV Wizard™ Plus SV 25 ml 150 prp 5 prp 10 col 25 col 100 prp 5 col 25 col 25 col 250 prp 50 rxn 400 prp 250 prp 50 prp kit 50 rxn 50 cap 10 prp 20 prp kit 20 col 20 col 20 col 50 col 20 col 250 prp 50 prp 120 prp 3 mg/ml 100 col 20 col 10 col 5 col 25 col 250 col 250 ml 50 col 50 col 250 col 50 rxn 250 rxn $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 176.74 101.38 324.78 151.62 359.77 1,327.82 187.51 177.64 122.02 253.00 58.32 538.31 329.26 70.88 70.88 70.88 48.45 107.23 107.23 107.23 71.38 103.54 71.38 133.35 103.54 768.88 174.95 139.43 171.36 83.87 56.77 90.35 86.36 111.78 99.95 94.35 59.20 65.09 222.20 42.56 189.73 FILM AND MEMBRANES ....................................................................................................... FM Mfgr. KOD KOD KOD FSH NEN KOD KOD FUJ FUJ Mfgr No V8701302 V8715187 V8294985 57144 NEF-978Y EB135-24 EB135-36 04-441-118 04-441-115 Code 2306 2307 2364 2118 2260 2382 2383 2325 2324 Item Description Biomax™ Film MR-1 8x10in Biomax™ Film MR-1 35x43cm Biomax™ Film MS-1 8x10in Chromatography Paper 3MM 46X57cm Colony/Plaque Screen™ w/orient. holes 137mm Elite II Slide Film 100 - 24 Exposures Elite II Slide Film 100 - 36 Exposures Fuji RX Film NIF 14x17in Fuji RX Film NIF 8x10in Unit 50 sht 50 sht 50 sht 100/pk 50/bx 1 roll 1 roll 100 sht 100 sht CC Price $ $ $ $ $ $ $ $ $ 138.93 300.20 118.70 85.24 231.21 6.76 9.13 305.28 111.69 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 63 NEN NEN NEN AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR AMR MIL MIL MIL RAD MSI MSI MSI MSI MSI MSI MIL MIL RAD RAD RAD INV INV MSI MSI POL POL NEN INV RAD RAD MIL MIL MIL MIL MIL MIL MIL NEF-986 NEF-976 NEF-984 RPN2020D RPN303D RPN137N RPN203N RPN2222N RPN303N RPN82N RPN87N RPN303E RPN137B RPN203B RPN2020B RPN303B RPN87B RPN2020F RPN1645 RPN1677L RPN2114K RPN3114K RPN1678K INYC13750 IPVH20200 IPVH00010 1620174 N00HN13750 NJ0HY13750 EP4HY408F0 EP4HY13750 EP4HYA0010 EP4HY00010 HATF08250 HATF13750 1620147 1620146 1620115 LC2000 LC2001 WP4HY08250 WP4HY00010 444159 444191 NEF-1002 LC2002 1620181 1620186 SCGPU02RE SCGPU05RE SCGPU11RE SCGPU01RE SCGP00525 SCGPS10RE SCGPT10RE 2250 2246 2249 2290 2333 2359 2139 2136 2137 2138 2358 2289 2357 2135 2292 2293 2356 2295 2308 4015 2286 2287 2143 2439 2214 2215 2440 2432 2150 2428 2157 2158 2159 2217 2216 2372 2370 2373 2441 2442 2164 2163 2322 2323 2253 2443 2371 2368 2271 2272 2273 2433 2339 2434 2435 Genescreen Plus® Hybridization Mem. 20x20cm 10/bx Genescreen Plus® Hybridization Mem. 30x45cm 5 sht Genescreen™ Hybridization Mem. 20x20cm 10/bx Hybond™ ECL™ 20x20cm 10 sht Hybond™ ECL™ 30cmx3m 1 roll Hybond™ Nylon 137mm 50/bx Hybond™ Nylon 20cmx3m 1 roll Hybond™ Nylon 22x22cm 10/bx Hybond™ Nylon 30cmx3m 1 roll Hybond™ Nylon 82mm 50/bx Hybond™ Nylon 87mm 50/bx Hybond™-C extra sup nitrocellulose 30cmx3m 1 roll Hybond™-N+ charged nylon 137mm 50/bx Hybond™-N+ charged nylon 20cmx3m 1 roll Hybond™-N+ charged nylon 20x20cm 10 sht Hybond™-N+ charged nylon 30cmx3m 1 roll Hybond™-N+ charged nylon 87mm 50/bx Hybond™-PVDF 20x20cm 10 sht Hypercassette™ (beige) 35x43cm 1 ea Hyperfilm MP Enveloped, 8x10in 50 sheets Hyperfilm™ ECL™ 8x10in 25 sht Hyperfilm™ ECL™ 8x10in 75 sht Hyperfilm™ MP 8x10in 75 sht Immobilon-NY+ 137mm DISC(Colony Plaque) 50/pk Immobilon™ P 20x20cm (Western Blot) 10/pk Immobilon™ P 26.5cmx3.75m (Western Blot) 1 roll Immun-Blot PVDF 10 sht Magna 137mm Notched 50/pk Magnagraph 137mm 50/pk Nitrobind 7x8.4 10/pk Nitrobind Pure 137mm 50/pk Nitrobind Pure 20cmx3m 1 roll Nitrobind Pure 30cmx3m 1 roll Nitrocellulose 82mm Colony & Plaque 50/pk Nitrocellulose 137mm Colony & Plaque 50/pk Nitrocellulose Membrane (.20u) 13.5x16.5cm 10 sht Nitrocellulose Membrane (.20u) 7x8.4cm 10 sht Nitrocellulose Membrane (.45u) 33cmx3m 1 roll Nitrocellulose Membrane (0.2 µm) Filter Paper Sandwich 20/pk Nitrocellulose Membrane (0.45 µm) Filter Paper Sandwich 20/pk Nitropure Supp 82mm 50/pk Nitropure Supp 30cmx3m 1 roll Polaroid Film Type 57 pk Polaroid Film Type 667 pk Polyscreen® PVDF 30cmx3m 1 roll PVDF Membrane/Filter Paper Sandwich 20/pk Sequi-Blot™ PVDF Membrane (.20u) 15x15cm 10 sht Sequi-Blot™ PVDF Membrane (.20u) 7x8.4cm 10 sht Stericup Filtr. & Stor.-PES 250ml .22µ 12/pk Stericup Filtr. & Stor.-PES 500ml .22µ 12/pk Stericup Filtr. & Stor.-PES 1000ml .22µ 12/pk Stericup Filtr. & Stor.-PES 150ml .22µ 12/pk Steriflip-GP Filter Unit 50µl 25/pk Steritop-GP Filtr 1000 ML 33 mm neck 0.22µm 12/pk Steritop-GP Filtr 1000 ML 45 mm neck 0.22µm 12/pk $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 150.24 218.43 149.17 136.93 168.86 182.30 195.74 157.94 214.22 89.05 99.13 236.06 231.86 200.78 195.74 223.46 126.85 178.51 204.16 144.28 79.55 206.34 206.34 156.48 101.81 125.90 55.49 107.32 125.41 23.91 104.99 99.17 146.88 49.63 101.81 126.00 46.47 171.57 104.21 104.21 50.75 140.31 47.95 24.07 181.14 101.89 99.19 64.34 42.64 66.82 95.45 33.09 80.25 65.54 65.54 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 64 MIL MIL MIL MIL MIL MIL KOD KOD KOD NEN NEN RAD SCGPS05RE SCGPT05RE SLGV025LS SLHVO25LS SLGS0250S SLHA0250S V1651678 V1651579 V1651454 NEF-586 NEF-596 1620192 2336 2335 2266 2264 2267 2265 2331 2332 2211 2388 2385 2362 Steritop-GP Filtr 500 ML 33 mm neck 0.22µm Steritop-GP Filtr 500 ML 45 mm neck 0.22µm Syringe fltr-Durapore PVDF/PVC .22µ Syringe fltr-Durapore PVDF/PVC .45µ Syringe fltr-mixed esters of cellulose .22µ Syringe fltr-mixed esters of cellulose .45µ X-Omat™ AR-2 interleaved film 35x43cm X-Omat™ AR-2 interleaved film 8x10in X-Omat™ AR-5 non-interleaved film 8x10in X-Omat™ Blue 5x7 X-Omat™ Blue 8x10in Zeta-Probe® GT Membrane15x15cm 12/pk 12/pk 50/pk 50/pk 50/pk 50/pk 50 sht 50 sht 50 sht 100/bx 100/bx 15 sht $ $ $ $ $ $ $ $ $ $ $ $ 41.36 41.36 70.63 66.49 64.27 62.36 353.46 182.70 144.05 57.53 135.27 180.51 IMMUNOLOGICAL REAGENTS .............................................................................................. IM Mfgr. AMR BMB AMR AMR INV CCS LTI LTI LTI COL PRO Mfgr No 27-4577-01 1583816 NA931 NA934 R96025 NA 15918-014 15920-010 19543-024 CB40066 S3841 Code 3186 3195 3143 3144 4636 3207 3111 3112 3116 3208 3203 Item Description Anti-GST Antibody Anti-HA (12CA5) Anti-Mouse Ig - HRP conjugated Anti-Rabbit Ig - HRP conjugated Anti-V5 Antibody myc 1 - 9E10 antibody Protein A Agarose Protein G Agarose Streptavidin Red 670™ TNF-a Western Blue Stabilized Substrate for AP Unit 0.5ml 200 µg 1 ml 1 ml 50 ul 0.5 mg 5 ml 5 ml 100 µg 10µg 1 ea CC Price $ 207.13 $ 202.38 $ 214.02 $ 165.42 $ 148.21 $ 20.00 $ 166.69 $ 253.01 $ 157.50 $ 225.88 $ 30.78 MOLECULAR BIOLOGY KITS ................................................................................................KT Mfgr. Mfgr No Code Item Description Unit CC Price BMB 1483188 5519 1st Strand cDNA Kit 30 rxn $ ENZ 42655-10 4630 BioArray High Yield RNA Transcript Labeling Kit 10 rxn $ ENZ 42655-20 4631 BioArray High Yield RNA Transcript Labeling Kit 20 rxn $ CLN 631312 6008 Cal Phos Mammalian Transfection Kit sm box $ RAD 5000112 5941 DC Protein Assay Kit II 1 kit $ QIA 69504 5473 DNeasy Tissue Kit 50 $ SIG XNAT-100RXN 4016 Extract n Amp Red 100 rxn $ AMR 27-9261-01 5314 First Strand cDNA Synthesis Kit 1 kit $ AMR 27-4590-01 5363 GST Detection Module 50 rxn $ AMR 27-4570-01 5362 GST Purifications Bulk Module 5 prps $ BMB 1585584 5281 High Prime DNA Labeling Kit 50 rxn $ BMB 1585592 5282 High Prime DNA Labeling mix for dCTP 50 rxn $ MLG 11452-018 5622 High Pure Maxi Kit 10rxn $ BMB 1796828 5520 High Pure PCR Template Prep EA $ QIA 12662 5471 Hispeed Plasmid Maxi Kit 10 prep $ QIA 12663 5472 Hispeed Plasmid Maxi Kit 25 prep $ LTI 18160-010 5133 Nick translation Kit 50 rxn $ INV K4900-01 6098 pcDNA3.1© Directional TOPO® Expression Kit (Top 10 Cells) 20 rxn $ STR 211190 4260 pCR-Script™ Amp Cloning Kit, (3 parts, -80,-20, RT) 1 kit $ STR 300385 5138 Prime-It® II Random Priming Kit 30 rxn $ 236.11 705.00 1,380.75 143.63 108.13 98.69 147.21 207.13 182.72 266.20 207.09 188.26 122.65 131.78 183.02 442.31 128.09 360.57 263.71 155.58 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 65 RAD RAD AMR STR LTI BMB MLG MLG MLG AMR USB USB RAD LTI INV PRO PRO PRO PRO PRO PRO INV INV INV INV INV INV 5000006 5000002 27-9558-01 200518 18428-011 1004760 11456-019 11458-015 11453-016 27-9240-01 70170 70140 1610449 11904-018 K2050-01 L4600 L5020 L4950 L4610 L1170 L5010 K4500-01 K4550-01 K4520-01 K4650-01 K4575-01 K2800-20 5937 5939 2167 5934 5227 5139 5625 5624 5623 4300 5278 5275 5360 5226 5389 5930 5931 5929 5928 5933 5932 6097 5397 5466 5398 5446 6096 Protein Assay Kit Dye Reagent Concentrate Protein Assay Kit II PureTaq RTG PCR Beads QuickChange™ Site-Directed Mutagenesis Kit Rad Prime Labeling Kit Random Primed DNA Labeling Kit Rapid Gel Extraction Kit Rapid PCR Purification Kit Rapid Plasmid Mini Prep Ready-To-Go DNA Labelling Beads (-dCTP) Sequenase™ PCR Product Seq. Kit Sequenase™ Quick Denature Seq. Kit Silver Stain Plus Kit Superscript First-Strand Synthesis System for RT-PCR TA Cloning Kit® with Dual Promoters (INVaF') TNT®-SP6 Coupled Transcription/Translation TNT®-SP6/T7 Coupled Transcription/Translation TNT®-T3 Coupled Transcription/Translation TNT®-T7 Coupled Transcription/Translation TNT®-T7 QUICK Coupled Transcription/Translation TNT®-T7/T3 Coupled Transcription/Translation TOPO TA Cloning® Kit (TOP10 Cells) TOPO TA Cloning® Kit (TOP10F' Cells) TOPO TA Cloning® Kit (with LTI 4622 - DH5a Cells) TOPO TA Cloning® Kit Dual Promoter (TOP10F' Cells) TOPO TA Cloning® Kit for Sequencing (Top 10 Cells) Zero Blunt® TOPO® PCR Cloning Kit (TOP 10 Cells) 1 kit 1 kit 0.5ML 1 kit 30 rxn 50 rxn 50rxn 50rxn 50rxn 1 kit 1 kit 1 kit kit 50 rxn 20 rxn 1 kit 1 kit 1 kit 1 kit 1 kit 1 kit 20 rxn 20 rxns 20 rxns 1 ea 20 rxn 20 rxn $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 69.70 81.32 142.55 475.24 128.88 209.44 54.28 54.28 48.83 102.38 262.92 184.10 116.69 235.30 289.93 291.15 300.09 312.11 293.37 297.05 314.95 322.35 308.67 319.57 322.99 334.39 338.20 MOLECULAR BIOLOGY PRODUCTS ...................................................................................MB Mfgr. Mfgr No Code Item Description Unit BMB 1204530 4267 ABTS® Buffer 125 ml BMB 1112422 4266 ABTS® Tablets (50mg) 20 tab NEB E8021L 4330 Amylose Resin 100 ml BMB 711454 4104 Bovine Serum Albumin (Nuclease free) 20 mg LTI 15561-020 4618 Bovine Serum Albumin, Acetylated 150 mg BMB 1213857 4233 Collagenase P 100 mg BMB 1202375 4126 DOTAP (Liposomal Transfection Reagent) 2 ml CLN 636831 6026 Express Hyb 250 ml BMB 3045595 5440 ExtremeGENE (Transfection Reagent) .4 ml PRO Q4100 4248 fmol® DNA Sequencing System 1 kit BMB 1815091 5421 FuGENE 6 (Nonliposomal Transfection Reagent) .4 ml BMB 1814443 5422 FuGENE 6 (Nonliposomal Transfection Reagent) 1 ml BMB 901393 4131 Glycogen 20 mg SIG G4877-10000iu 6067 GONADOTROPIN FROM PREGNANT MARES' SERUM 10000iu LTI 15279-011 4135 Human Cot-1® DNA 500 µg LTI 11668-019 6031 LipofectAMINE 2000 1.5ml LTI 11668-027 6032 LipofectAMINE 2000 0.75ml LTI 18324-012 4181 Lipofectamine™ 1 ml LTI 18324-020 4582 Lipofectamine™ 4 x 1 ml LTI 18324-111 4624 Lipofectamine™ .5 ml LTI 18292-011 4138 Lipofectin® Reagent 1 ml NEB S1404S 4331 m7GpppG 5'G RNA Cap Structure 25u LTI 17502-048 4620 N2 Supplement 5 ml CC Price $ 23.53 $ 40.79 $ 220.61 $ 49.42 $ 98.67 $ 50.20 $ 242.39 $ 148.63 $ 84.72 $ 143.73 $ 125.51 $ 250.23 $ 77.66 $ 339.13 $ 108.53 $ 273.82 $ 166.04 $ 165.28 $ 543.86 $ 98.67 $ 125.82 $ 110.31 $ 38.68 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 66 LTI PRO LTI NEB PRO PRO PRO LTI BMB CCS CCS CCS CCS CCS CCS CCS CCS CCS CCS BMB STR PRO AMR BMB AMR AMR FSH PRO PRO PRO PRO STR CLN CLN PRO LTI LTI CLN CLN INV 18418-012 C1101 12252-011 N3032S A1360 A1380 A3600 11514-015 108812 4463 4464 4465 4466 4190 4191 4194 4192 4193 4324 3335399 201220 L4960 27-2166-01 1034731 RPN1635 27-0816-01 2123621 N2515 N2511 N2115 N2111 201190 635501 635506 E2381 15596-026 15401-011 630440 630439 R250-01 6125 4268 4009 4254 5942 5943 4294 6034 4152 4463 4464 4465 4466 4190 4191 4194 4192 4193 4524 5525 4551 4169 4183 4170 4239 5357 4201 4176 4177 4174 4175 4552 6027 6028 5944 4182 4180 6030 6029 5417 Oligo (dT) 12-18 Primer Oligo(dT) 15 primer Oligofectamine Reagent pBR322/Msp I Digest pGEM®-T Easy I pGEM®-T Easy II pGEM®-T Vector I Plus Reagent Poly d(I-C) Primer gt10 forward Primer gt10 reverse Primer gt11 forward Primer gt11 reverse Primer M13/pUC Forward 23mer Primer M13/pUC Reverse 23mer Primer Sp6 20mer Primer T3 20mer Primer T7 20mer Primer T7 Term Protector RNase Inhibitor QuickHyb® Rapid Hybridization Buffer Rabbit Reticulocyte Lysate Random Primers (N)6 Random Primers (N)6 RapidHyb Hybridization™ Buffer RNAguard Ribonuclease Inhibitor RNase AWAY RNasin® Recombinant Ribonuclease Inhibitor RNasin® Recombinant Ribonuclease Inhibitor RNasin® Ribonuclease Inhibitor RNasin® Ribonuclease Inhibitor Salmon Sperm DNA Talon Metal Affinity Resin Talon Superflow Metal Affinity Resin Tfx-10 Transfection Reagent TRIzol Reagent Yeast tRNA Yeastmaker Carrier DNA Yeastmaker Yeast Transformation System Zeocin 25 UG 20 µg 1 ML 50 µg 1 kit 1 kit 20u .85 ml 10 Å260u 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg EA 250 ml 1 ml 50 Å260u 50 Å260u 125 ml 5000u 8.4 oz 10000u 2500u 10000u 2500u 10 x 1ml 10 ml 25 ml 1 ea 100 ml 25 mg ea kit 1g $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 45.40 33.28 188.12 40.44 89.93 151.91 74.41 87.73 66.68 20.00 20.00 20.00 20.00 20.00 20.00 20.00 20.00 20.00 20.00 64.32 154.95 94.41 117.35 111.39 65.37 55.13 17.05 173.32 61.49 163.43 64.86 156.16 69.64 194.59 143.21 78.80 44.91 199.23 203.86 175.07 PROTEIN AND MOLECULAR WEIGHT MARKERS .................................……………………………............................MR Mfgr. LTI LTI NEB LTI PRO LTI PRO LTI NEB PRO Mfgr No 15615-016 15615-024 N3232S 10381-010 PRG6941 10787-018 G4471 15628-019 N3231S G2101 Code 4123 4221 4584 4302 4588 4625 4585 4101 4583 4455 Item Description 1 kb DNA Ladder 1 kb DNA Ladder 1 kb DNA Ladder 1 kb DNA Ladder - RTS - ready load 1 kb DNA Step Ladder 1 kb Plus DNA Ladder 10 bp DNA Step Ladder 100 bp DNA Ladder 100 bp DNA Ladder 100 bp DNA Ladder Unit 250 µg 1000 µg 100ug 100 app 300 lns 250 µg 50 µl 50 µg 50ug 250 µl CC Price $ 93.19 $ 309.87 $ 40.44 $ 53.13 $ 58.57 $ 84.88 $ 57.08 $ 41.70 $ 40.44 $ 56.38 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 67 PRO LTI LTI LTI PRO LTI AMR LTI PRO LTI BMB BMB BMB BMB BMB BMB BMB RAD NEB LTI NEB PRO LTI INV INV INV INV LTI NEB NEB PRO LTI RAD NEB RAD RAD RAD AMR AMR LTI LTI INV INV INV G6951 10380-012 10068-013 15613-011 G6961 10597-011 27-4005-01 10416-014 G4521 10748-010 1418009 1449460 821705 1062590 1336045 1498037 1721933 1610324 N3011S 15612-013 N3012S G1711 10382-018 LC5600 LC5602 LC5677 LC5725 15611-015 N3026L N3026S G1761 10383-016 161-0374 P7708S 1610318 1610309 1610305 RPN756 RPN755 12308-011 15620-016 LC5925 LC5625 LC6060 4586 4304 4196 4102 4587 4498 4484 4499 4590 4619 4480 4231 4229 4230 4545 4232 4454 4515 4252 4136 4137 4486 4307 5182 4014 4596 4597 4151 4250 4251 4198 4305 4633 4546 4483 4482 4481 4234 4235 4632 4172 4599 4598 4600 100 bp DNA Step Ladder 100 bp Ladder - RTS - ready load 100 bp-2kb DNA Mass Ladder 123 Base Pair Ladder 200 bp DNA Step Ladder 25 bp DNA Ladder 50 bp DNA Ladder 50 bp DNA Ladder 50 bp DNA Ladder BENCHMARK™ Prestained Protein Ladder DNA Marker IV (74bp - 19329bp) DNA Marker IX (72-1353bp) DNA Marker V (8-587bp) DNA Marker VI (154-2176bp) DNA Marker VIII (19bp - 1114bp) DNA Marker X (75-12216bp) DNA Marker XIV (100 - 1500 bp) 100 bp ladder Kaleidoscope Prestained Standards Lambda DNA Lambda DNA/HindIII Fragment Lambda DNA/HindIII Fragment Lambda DNA/HindIII Fragment Lambda DNA/HindIII Fragment - RTS - ready load MagicMark Western Protein Standard MagicMark XP Mark12™ Wide Range Molecular Weight Standard MultiMark® Multi-Colored Standard Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment - RTS - ready load Precision Plus Protein Dual Color Standard Prestained Protein Molecular Weight Marker Prestained Standards Broad Prestained Standards High Prestained Standards Low Rainbow Marker™ (14300-250000) Rainbow Marker™ (2350-46000) Ready Load 1KB Plus DNA Ladder RNA Ladder 0.24-9.5 kb SeeBlue® Plus2 Pre-Stained Standard SeeBlue® Pre-Stained Standard SimplyBlue™ SafeStain 100 µl 100 app 200 µl 100 µg 100 µl 50 µg 200 µg 50 µg 100 lns 2 x 250u 50 µg 50 µg 50 µg 50 µg 50 µg 100 µg 50 µg 500 µl 250 µg 500 µg 150 µg 100µg 100 app 250ul 250ul 1.0 ml 500 µl 40 µg 250u 50u 50 µg 100 app 50 app 175 lns 500 µl 500 µl 500 µl 250 µl 250 µl 100 rxn 75 µg 500 µl 500 µl 1L $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 75.92 64.05 64.41 67.10 78.51 71.58 154.36 71.58 65.48 78.71 83.15 105.11 83.93 87.86 100.41 87.86 74.52 89.36 40.44 90.41 40.44 24.45 44.63 177.71 178.10 92.63 106.52 54.30 161.78 40.44 65.24 83.60 93.76 62.51 80.42 75.96 68.81 101.12 101.12 73.79 80.29 99.11 99.11 79.66 MISCELLANEOUS PRODUCTS ............................................................................................. MS Mfgr. FSH AMR FSH FSH FSH FSH Mfgr No 121311 NAMP100 212363 212362A 212368 212361 Code 6023 4296 4222 4228 4226 4227 Item Description Aluminum Foil (18” x 50ft) Amplify™ ART® 10 aerosol resistant pipet tip (racked) ART® 1000E aerosol resistant pipet tip (racked) ART® 100E aerosol resistant pipet tip (racked) ART® 200 aerosol resistant pipet tip (racked) Unit ea 1L 960/pk 800/pk 960/pk 960/pk CC Price $ 8.53 $ 143.86 $ 57.00 $ 57.00 $ 57.00 $ 57.00 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 68 FSH FSH DEN INV INV INV RAD RAD RAD TKR TKR QIA QIA DEN DEN DEN DEN DEN RAI RAI RAI RAI RAI RAI RAI RAI RAI RAI RAI RAI DEN DEN DEN DEN DEN DEN FSH 212365 212364 R6651 P410-50 P450-50 P460-50 1652089 1652086 1652088 TKR-1301 C1201-CL 2003482 2003481 P-1125 P-1123 P-1096-FR P-1121 P-1122 L-10 L-2 L-100 L-200 L-20 L-1000 P-2 P-10 P-1000 P-100 P-20 P-200 P3950-10 P3950-100 P3950-1000 P3950-20 P3950-200 P3950-5000 15610 4224 4223 4462 4519 4520 4521 5950 4517 4518 4621 4497 3345 3344 3327 3329 3325 3326 3328 6037 6036 6039 6040 6038 6035 5955 5956 5954 5952 5951 5953 4456 4458 4460 4457 4459 4461 6024 ART® 20E aerosol resistant pipet tip (racked) ART® 20P aerosol resistant pipet tip (racked) Chillete personal tube cooler -12 tubes at -20C Electroporation Cuvettes Electroporation Cuvettes Electroporation Cuvettes Gene Pulser® Cuvettes 0.1cm Gene Pulser® Cuvettes 0.2cm Gene Pulser® Cuvettes 0.4cm Mini-Centrifuge Mini-Centrifuge 6 x 1.5 ml Oligo CD - $1,000 Oligo CD - $500 P-100 Filter Pipet Tips P-1000 Filter Pipet Tips P2/P10 Eatra Long Filter Pipet Tip P-20 Filter Pipet Tips P-200 Filter Pipet Tips Pipete-Lite Magnetic Assist Pipette .5-10µl Pipete-Lite Magnetic Assist Pipette 0.1- 2µl Pipete-Lite Magnetic Assist Pipette 10-100µl Pipete-Lite Magnetic Assist Pipette 20-200µl Pipete-Lite Magnetic Assist Pipette 2-20µl Pipet-Lite Magnetic Assist Pipette, 100-1000ul Pipetman 0.1-2.0µl Pipetman 0.5-10µl Pipetman 100-1000µl Pipetman 10-100µl Pipetman 2-20µl Pipetman 50-200µl Pipet-mate Digital Pipette w/ tip ejector 10µl Pipet-mate Digital Pipette w/ tip ejector 100µl Pipet-mate Digital Pipette w/ tip ejector 1000µl Pipet-mate Digital Pipette w/ tip ejector 20µl Pipet-mate Digital Pipette w/ tip ejector 200µl Pipet-mate Digital Pipette w/ tip ejector 5000µl Wrapping Film (18in x 1000ft) 1000/pk 960/pk 1 ea 0.1cm 0.2cm 0.4cm 50 pk 50 pk 50 pk ea 1 ea EA EA EA EA EA EA EA 1 ea 1 ea 1 ea 1 ea 1 ea ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea ea $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 56.98 57.00 28.60 115.26 115.26 115.26 116.69 124.21 116.69 250.00 185.59 1,000.00 500.00 47.30 47.30 47.30 47.30 47.30 227.60 227.60 199.60 199.60 199.60 199.60 266.92 266.92 234.45 234.45 234.45 234.45 158.95 152.01 152.01 152.01 152.01 159.29 18.66 NON-RADIOACTIVE KITS AND REAGENTS .....................................................................………………………... NR Mfgr. Mfgr No Code Item Description Unit CC Price BMB 1096176 3154 Blocking Reagent 50 g NEN NEL-601 5979 CDP-Star Nucleic Acid Chemiluminescence Reagent (Ready to Use) 1 kit BMB 1644807 3181 Cell Proliferation Kit (WST-1) ea BMB 1500708 3123 Chemiluminescent Western Blot Kit 1000cm2 BMB 1585614 3178 DIG/Genius High Prime DNA Labeling and Detection Starter Kit II 1 kit BMB 1277073 3175 Digoxigenin RNA Labeling Mix 20 rxn BMB 1209256 3151 Digoxigenin-11-UTP 250 nmol AMR RPN2132 5415 ECL™ Plus Western Blotting Detection Reagents 1 ea AMR RPN2108 3117 ECL™ Western Blot Analysis Starter System 1 kit AMR RPN2109 3118 ECL™ Western Blotting Detection Reagents 1000cm2 AMR RPN2106 3119 ECL™ Western Blotting Detection Reagents 4000cm2 AMR RPN2209 3142 ECL™ Western Blotting Detection Reagents 2000cm2 BMB 1175025 3130 GENIUS™ 4 RNA Labeling Kit 2x10rxn University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 69 $ $ $ $ $ $ $ $ $ $ $ $ $ 79.23 134.25 278.47 76.87 192.18 92.56 123.94 181.69 131.75 103.33 225.60 162.74 311.42 BMB PRO PRO PRO BMB BMB NEN NEN NEN NEN BMB BMB BMB 1684795 E4030 E1501 E4530 2015200 2015196 NEL-201 NEL-103 NEL-104 NEL-105 1093274 1767305 1767291 3173 3185 3212 3211 6041 6042 5201 3120 3121 3122 3132 3180 3179 In Situ Cell Death Detection Kit - Fluorescein Luciferase Assay System Luciferase Assay System (10 pk) quote#5075 Luciferase Assay System–10 pack Lumi Light Lumi Light + Renaissance™ Nucleic Acid Chemilum Rgt Kit Renaissance™ Western Blot Chemilum Rgt Renaissance™ Western Blot Chemilum Rgt Renaissance™ Western Blot Chemilum Rgt Sheep Anti-digoxigenin-AP TUNEL Enzyme (optimized terminal transferase) TUNEL Label (FITC dUTP + dNTPs) 50 rxn 100 assays 1000 assays 1000 assays 1 kit 1 kit 1 kit 1000cm2 2500cm2 5000cm2 150u 1 ea 1 ea $ $ $ $ $ $ $ $ $ $ $ $ $ OTHER ENZYMES .................................................................................................................. OE Mfgr. BMB NEB BMB USB BMB PRO PRO BMB NEB PRO PRO NEB LTI AMR LTI LTI BMB BMB PRO LTI LTI AMR PRO PRO BMB LTI NEB PRO AMR LTI PRO AMR AMR LTI PRO BMB BMB Mfgr No 1417215 M0392S 1758250 70092Y 109118 M5108 M5101 713023 M0290S M1821 M2051 M0209S 18010-025 27-0514-01 18047-019 18068-015 104159 776785 M6101 18052-019 10480-010 E70073Y M1811 M2181 1008404 18012-021 M0210S M2201 27-0925-01 28025-013 M1701 E2420Y RPN1633 18021-014 M4281 1119915 109169 Code 5101 5307 5374 5102 5103 5105 5106 5108 5110 5256 5204 5112 5183 5373 5113 5225 5320 5114 5144 5454 5475 5287 5262 5366 5125 5120 5122 5123 5128 5127 5367 5131 5375 5141 5205 5143 5386 Item Description Agarase Agarase Alkaline Phosphatase (shrimp) Alkaline Phosphatase (shrimp) AMV Reverse Transcriptase AMV Reverse Transcriptase AMV Reverse Transcriptase Calf Intestinal Alkaline Phosphatase Calf Intestinal Alkaline Phosphatase Calf Intestinal Alkaline Phosphatase DNA Polymerase I DNA Polymerase I from E.coli DNA Polymerase I (E Coli) DNase 1 DNase I DNase I - Amplification Grade DNase I - Grade II DNase I - RNase Free DNase RQ1 RNase-free E. Coli DNA Ligase Elongase Enzyme Mix Exonuclease I Exonuclease III Klenow - Exo Minus Klenow - Labeling grade Klenow - Sequencing Grade Klenow - Sequencing Grade Klenow Fragment Maloney-MLV Reverse Transcriptase Maloney-MLV Reverse Transcriptase Maloney-MLV Reverse Transcripts Mung Bean Nuclease Rediprime™ kit for dCTP Ribonuclease H Ribonuclease H RNase - DNase free Rnase A Unit 100u 100u 1000u 500u 1000u 1000u 300u 1000u 1000u 1000u 500u 500u 1000U 1000u 20000u 100u 100 mg 10000u 1000u 100u 100 rxn 2500u 5000u 100u 100u 100u 200u 150u 200u 40000u 10000u 3000u 30 rxn 30u 50u 500 µg 100 mg CC Price $ 175.71 $ 44.12 $ 69.03 $ 85.48 $ 200.81 $ 131.21 $ 50.41 $ 47.07 $ 40.44 $ 35.58 $ 66.72 $ 40.44 $ 113.64 $ 63.79 $ 184.22 $ 56.95 $ 36.08 $ 109.04 $ 22.19 $ 23.20 $ 100.62 $ 81.94 $ 45.70 $ 27.43 $ 47.85 $ 39.22 $ 36.77 $ 54.87 $ 55.13 $ 141.01 $ 31.88 $ 118.92 $ 186.66 $ 73.88 $ 97.86 $ 57.26 $ 57.26 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 70 266.70 46.84 308.22 412.41 142.77 247.09 131.06 77.79 140.65 198.18 127.86 92.56 69.03 AMR BMB PRO USB USB USB USB USB AMR LTI PRO EPI LTI LTI LTI LTI LTI PRO BMB LTI LTI NEB NEB PRO NEB BMB LTI LTI NEB PRO BMB BMB LTI NEB NEB PRO AMR PRO AMR LTI PRO AMR BMB E70054Y 109193 M5761 70710 70754 70721 70724 70775z 27-0808-01 18018-010 P1085 SL910K 11917010 18080044 12574-026 18064-014 18053-017 P2083 481220 15224-017 15224-025 M0202S M0202L M1801 M0202T 1004786 18005-017 18005-025 M0203S M4211 1635379 709557 18004-010 M0201S M0201L M4101 27-0736-01 P2075 E2230Z 10533-065 M1871 27-0846-01 109541 5289 5388 5207 5147 5148 5150 5151 5239 5371 5156 5158 5322 5460 5478 5476 5159 5160 5161 5163 5164 5229 5165 5217 5208 5166 5167 5168 5464 5169 5209 5259 5171 5172 5173 5218 5175 5174 5179 5233 5194 5210 5368 5387 RNase H RNase T1 S1 Nuclease Sequenase™ dGTP labeling mix Sequenase™ dGTP Nucleotide Kit Sequenase™ Reagent Kit Sequenase™ Stop Solution Sequenase™ v 2.0 SP6 RNA Polymerase Sp6 RNA Polymerase Sp6 RNA Polymerase SP6 RNA Polymerase (25u/µl) Superscript Custom Choice Kit Superscript III Reverse Transcriptase SuperScript III, 1 step w/Platinum Taq 100 Superscript™ II Reverse Transcriptase Superscript™ Reverse Transcriptase T3 RNA Polymerase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase HC T4 DNA Polymerase T4 DNA Polymerase T4 DNA Polymerase T4 DNA Polymerase T4 DNA Polymerase T4 DNA Rapid Ligation Kit (5 minutes) T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase FPLCpure T7 RNA Polymerase Terminal Deoxy Transferase (TdT) Terminal Deoxy Transferase (TdT) Terminal Deoxy Transferase (TdT) Thrombin Protease tRNA 250u 100000u 10000u 100 µl 1 kit 1 kit 1.2 ml 1000u 3000u 500u 1000u 10000u 10 rxn 10000u 100 rxn 10000u 10000u 1000u 100u 100u 500u 20000u 100000u 100u 20000u 100u 50u 250u 150u 100u 40 rxn 200u 200u 500u 2500u 100u 200u 1000u 2500u 500u 300u 500u 100 mg $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 51.19 33.73 19.27 25.20 119.41 90.93 17.46 417.91 152.79 48.07 94.73 102.02 656.60 158.13 351.55 149.91 154.90 22.97 40.79 26.09 115.12 44.12 176.48 22.98 44.12 73.74 31.89 103.79 40.44 48.19 104.33 160.81 50.03 36.77 147.07 27.43 73.24 30.39 345.75 108.42 39.29 62.22 92.56 PCR REAGENTS AND NUCLEOTIDES ............................................................................………………….. PC Mfgr. LTI LTI CLN CLN CLN PEC PEC Mfgr No 18427-088 18427013 639102 639105 639505 N808-0246 N808-0240 Code 4010 5468 5985 5987 6051 5993 5382 Item Description 10 mM dNTP Mix 10mM dNTP Mix Advantage cDNA PCR Kit Advantage cDNA Plolymerase Mix Advantage RT for PCR AmpliTaq Gold™ w/ buffer I - 1000u AmpliTaq Gold™ w/ buffer I - 250u Unit 1 ML 100 µl 1 ea 1 ea ea 1000u 250u CC Price $ 216.22 $ 28.61 $ 139.34 $ 254.83 $ 241.50 $ 284.65 $ 86.08 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 71 PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC PEC AMR BMB PEC NEN NEN BMB AMR BMB PRO AMR BMB AMR BMB PRO AMR BMB AMR BMB USB USB USB AMR USB SIG PRO AMR BMB AMR BMB AMR AMR DEN AMR PRO BMB N808-0244 N808-0248 N808-0242 N808-0247 N808-0241 N808-0245 N808-0249 N808-0243 N808-0171 N808-0152 N808-0160 N808-0155 N808-0166 N808-0172 N808-0161 N808-0156 N808-0167 N808-0153 N808-0150 N808-0100 27-2056-01 1140965 4303573 NEL542 NEL999 1093070 27-2066-01 1140922 U1201 27-2050-01 1051440 27-2050-02 1934511 U1221 27-2060-01 1051458 27-2060-02 1934520 70714 70716 70718 27-2045-01 70720 D7295-.5ml U1211 27-2070-01 1051466 27-2070-02 1934538 27-2094-01 27-2035-02 CB-4420-2 27-2035-01 U1240 1969064 5997 5999 5995 5994 5383 5998 6000 5996 5263 5264 5212 5265 5266 5267 5213 5268 5269 5270 5251 5250 5343 4103 5407 5455 5456 3133 5345 4105 4241 5347 4106 5449 6044 4242 5348 4108 5450 6045 4110 4111 4112 5352 4114 5411 4243 5349 4115 5451 6046 5336 5353 3307 4125 4240 6043 AmpliTaq Gold™ w/ buffer I-12 pac (250u) AmpliTaq Gold™ w/ buffer I-5 pac (1000u) AmpliTaq Gold™ w/ buffer I-6 pac (250u) AmpliTaq Gold™ w/ buffer II - 1000u AmpliTaq Gold™ w/ buffer II - 250u AmpliTaq Gold™ w/ buffer II-12 pac (250u) AmpliTaq Gold™ w/ buffer II-5 pac (1000u) AmpliTaq Gold™ w/ buffer II-6 pac (250u) AmpliTaq® w/ buffer I - 1000u AmpliTaq® w/ buffer I - 12 paq (250u) AmpliTaq® w/ buffer I - 250u AmpliTaq® w/ buffer I - 5 paq (1000u) AmpliTaq® w/ buffer I - 6 paq (250u) AmpliTaq® w/ buffer II - 1000u AmpliTaq® w/ buffer II - 250u AmpliTaq® w/ buffer II - 5 paq (1000u) AmpliTaq® w/ buffer II - 6 paq (250u) AmpliTaq® w/ buffer II- 12 paq (250u) AmpliWax® PCR Gem 50 AmpliWax® PCR Gem 100 ATP 0.1M ATP 0.1M BigDye™ Terminator Kit Biotin 11 CTP Biotin 16 UTP Biotin-16-dUTP CTP 0.1M CTP 0.1M dATP (100mM) dATP 0.1M dATP 0.1M dATP 100mM solution DATP PCR Grade dCTP (100mM) dCTP 0.1M dCTP 0.1M dCTP 100mM solution DCTP PCR Grade ddATP Termination Mix ddCTP Termination Mix ddGTP Termination Mix ddNTP Set ddTTP Termination Mix Deoxynucleotide Mix dGTP (100mM) dGTP 0.1M dGTP 0.1M dGTP 100mM solution DGTP PCR Grade DNA Polymerization Mix dNTP Set 4x100umol dNTP Set (4x25umol) 100mM dNTP Set 0.1M dNTP's (bulk) (100mM) 4x40umoles dNTP's PCR Grade 3000u 5000u 1500u 1000u 250u 3000u 5000u 1500u 1000u 3000u 250u 5000u 1500u 1000u 250u 5000u 1500u 3000u 200 gem 200 gem 25 µmol 40 µmol 24 rxn 250nm 250nm 50 nmol 25 µmol 40 µmol 40 µmol 25 µmol 25 µmol 100 µm ea 40 µmol 25 µmol 25 µmol 100 µm ea 250 µl 250 µl 250 µl 4x0.5µmol 250 µl 1 vl 40 µmol 25 µmol 25 µmol 100 µm ea 4x10umol 4x100µmol EA 25 µmol 1 set ea $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 1,032.97 1,413.18 516.49 284.65 86.08 1,032.97 1,413.18 516.49 288.37 900.62 75.05 1,431.72 450.32 288.37 75.05 1,431.72 450.32 900.62 73.50 73.50 48.04 63.54 192.37 187.07 186.03 231.41 48.04 63.54 61.74 47.25 47.07 146.49 47.07 61.74 47.25 47.07 146.49 47.07 25.20 25.20 25.20 107.11 25.20 25.61 61.74 47.25 47.07 146.49 47.07 83.48 519.80 130.45 159.88 193.18 165.51 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 72 PRO AMR BMB AMR BMB BMB BMB BMB BMB BMB BMB BMB BMB BMB PEC PEC PEC PEC PEC PEC PEC PEC AMR STR STR QIA QIA FSH FSH FSH SIG BMB BMB FSH FSH FSH FSH AMR USB BMB PRO BMB BMB STR STR STR STR STR STR LTI LTI LTI LTI LTI LTI U1231 27-2080-01 1051482 27-2080-02 1934546 1732641 1732650 1759078 1681834 1759060 1681842 2032902 2032929 2140306 N808-0260 4303442 4303441 N808-0009 N808-0007 N808-0179 N808-0143 N808-0193 27-2076-01 600262 600260 203205 203203 E954140156 E954140164 E954140172 D5062-500u 2239264 3003230 E0032002250 E0032002404 E0032002501 E0032002552 27-2025-01 77102 1696513 PRG3161 1581295 1636138 600153 600154 600380 600250 600252 600320 11708-013 11708-021 11708-039 10790020 10966-018 10966-026 4244 5350 4127 5452 6047 5325 6011 6018 5274 6003 6012 5442 5443 5444 5404 5406 5405 5191 5247 5253 5202 5248 5344 6090 6089 6100 6099 5514 5515 5516 5410 5523 5524 5513 5510 5511 5512 5342 5241 5327 4199 4197 4289 5400 5401 4008 5402 5403 5447 6052 6053 6054 5459 6013 6014 dTTP (100mM) dTTP 0.1M dTTP 0.1M dTTP 100mM solution DTTP PCR Grade Expand™ High Fidelity PCR System Expand™ High Fidelity PCR System Expand™ High Fidelity PCR System Expand™ Long Template PCR System Expand™ Long Template PCR System Expand™ Long Template PCR System FastStart Taq DNA Polymerase FastStart Taq DNA Polymerase GC Rich Polymerase GeneAmp® dNTP blend 10mM GeneAmp® dNTP blend 10mM-12 paq GeneAmp® dNTP blend 10mM-6 paq GeneAmp® Core Kit GeneAmp® dNTP Set -10mM (4x3.2umol) GeneAmp® EZ rTth RNA PCR Kit GeneAmp® RNA PCR Core Kit GeneAmp® XL PCR Kit GTP 0.1M Herculase Enhanced DNA Polymerase Herculase Enhanced DNA Polymerase Hot Star Taq (1000) with Q solution Hot Star Taq (250) with Q solution HotMaster Taq w/ buffer HotMaster Taq w/ buffer HotMaster Taq w/ buffer Klentaq LA DNA Polymerase LC FastStart DNA Mstr Sybrgreen LC FastStart DNA Mstr Sybrgreen MasterMix MasterTaq Kit MasterTaq Kit MasterTaq Kit NTP Set 0.1M Nucleix-dATP set aq. (200uM) 0.1M PCR Cleanup Kit PCR Markers PCR Nucleotide Mix PCR Optimization Kit (for optimal pH & Mg) Pfu DNA Polymerase Pfu DNA Polymerase Pfu Ultra High Fidelity PfuTurbo™ DNA Polymerase PfuTurbo™ DNA Polymerase PfuTurbo™ HotStart DNA Polymerase Platinum Pfx DNA Polymerase Platinum Pfx DNA Polymerase Platinum Pfx DNA Polymerase Platinum Supermix Hi Fi Platinum Taq DNA Polymerase - 100u Platinum Taq DNA Polymerase - 250u 40 µmol 25 µmol 25 µmol 100 µm ea 100u 500u 10x250u 100u 10x250u 500u 100u 500u 50 rxn 1 ml 12 ml 6 ml 100 rxn 1 set 100 rxn 100 rxn 100 rxn 25 µmol 500u 100u (4x250u) 250u 100u 250u 1000u 500u EA EA 152u 100u 250u 500u 4x25µmol 200uM 100 rxn 250 µl 100 rxn 1 kit 100u 500u 100 u 100u 500u 100u 100 u 250 u 500 u 100 rxns 100u 250u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 61.74 42.53 47.07 146.49 47.07 69.03 324.75 1,455.11 73.74 1,546.10 342.01 59.62 283.18 146.69 29.40 352.80 176.41 164.21 36.50 355.64 237.21 375.40 48.04 295.62 68.03 261.24 92.42 31.95 74.55 266.25 162.58 828.35 207.09 39.94 22.37 55.91 106.50 159.88 148.99 111.39 54.12 49.42 123.15 85.98 345.80 102.99 93.54 382.65 104.88 81.62 185.54 350.35 121.70 46.05 110.99 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 73 LTI LTI QIA SIG PEC PEC PEC LTI LTI PEC PEC SIG QIA QIA SIG PRO BMB BMB BMB LTI LTI PRO PRO FSH FSH SIG BMB PEC PEC SIG BMB BMB BMB BMB BMB AMR NEB NEB 10966-034 11304-011 204143 D4309-250u N808-0178 N808-0098 N808-0180 10928-034 18064-071 4309155 4312704 S4438-100rxn 201203 201207 D1806-250u M1868 1146165 NA 1435094 18038-018 18038-042 M1861 M1865 FB600010 FB600045 D4545-250u 1578553 4304437 4305719 T4809 3186172 3186202 3186199 1855476 1888382 27-2086-01 M0257S M0254S 6015 5469 4634 6055 5255 5254 5249 5467 5465 4628 4629 5477 6001 6025 5408 5982 5185 5198 6002 5187 5188 5189 5258 5309 5311 5409 5199 4626 4627 5414 5461 5462 5463 5885 5988 5346 5257 5195 Platinum Taq DNA Polymerase - 500u Platinum Taq High Fidelity QuantiTect SYBR Green PCR Kit Redtaq polymerase rTth DNA Polymerase & EZ Buffer Pack rTth DNA Polymerase with Buffer rTth DNA Polymerase-XL w/XL Buffer Pack SuperScript One Step RT-PCR with Platinum Taq Superscript™ II Reverse Transcriptase Sybr Green PCR Master Mix Sybr Green PCR Master Mix SYBR Green qPCR Taq DNA Polymerase Taq DNA Polymerase Taq DNA Polymerase Taq DNA Polymerase Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer A Taq DNA Polymerase w/ buffer B Taq DNA Polymerase w/10X Buffer Taq PCR Core Kit TaqMan Universal Master Mix TaqMan Universal Master Mix Taqstart Antibody Tgo DNA Polymerase Tgo DNA Polymerase Tgo DNA Polymerase TITAN™ One-Tube RT-PCR System TITAN™ One-Tube RT-PCR System UTP 0.1M Vent® (exo-) Polymerase Vent® DNA Polymerase 500u 100u 200 rxn 250 unit 100 rxn 100 rxn 100 rxn 25 rxn 4 x 10000u pk 10 pk 100 rxn 250u 10x250u 250u 2500u 100u 250u 5000u 100u 500u 100u 500u 100u 100u 250u 100 rxn 200 rxn 2000 rxn 200 tst 50 rxns 100 rxns 250 rxns 100 rxn 25 rxn 25 µmol 200u 200u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 200.72 102.47 296.07 84.14 229.19 229.19 190.34 117.78 599.63 273.52 2,679.00 121.46 50.37 1,044.95 78.69 433.71 53.34 65.00 1,255.08 29.01 115.86 24.33 82.75 149.17 149.17 80.27 185.91 310.00 3,035.00 183.51 93.35 156.89 327.11 336.52 105.90 48.04 58.83 58.83 PRECAST GELS ......................................................................................................... PG Mfgr. INV INV INV INV INV INV INV INV INV INV INV INV Mfgr No EA0375 EA03752 EA0378 EA0355 EA0358 NP0342 NP0335 NP0301 NP0302 NP0303 NP0341 NP0343 Code 6068 6069 6070 6071 6072 4013 4012 6073 6074 6075 6076 6077 Item Description 3-8% NuPAGE® Tris-Acetate Gels 1.0 mm 10 well 3-8% NuPAGE® Tris-Acetate Gels 1.0 mm 12 well 3-8% NuPAGE® Tris-Acetate Gels 1.5 mm 10 well 7% NuPAGE® Tris-Acetate Gels 1.0 mm 10 well 7% NuPAGE® Tris-Acetate Gels 1.5 mm 10 well NuPAGE 12% Bis-Tris Gels 1.0mm 12 well NuPAGE 4-12% Bis-Tris Gels 1.5mm 10 well NuPAGE® 10% Bis-Tris Gels 1.0 mm 10 well NuPAGE® 10% Bis-Tris Gels 1.0 mm 12 well NuPAGE® 10% Bis-Tris Gels 1.0 mm 15 well NuPAGE® 12% Bis-Tris Gels 1.0 mm 10 well NuPAGE® 12% Bis-Tris Gels 1.0 mm 15 well Unit ea ea ea ea ea ea EA ea ea ea ea ea CC Price $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 $ 9.73 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 74 INV INV INV RAD RAD RAD RAD NP0321 NP0322 NP0323 1611101 1611155 1611104 1611158 6078 6079 6080 3340 3342 3341 3343 NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 10 well NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 12 well NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 15 well ReadyGel, 10% Tris-Hcl, 10 well ReadyGel, 10% Tris-Hcl, 50ul ReadyGel, 4-15% Tris-Hcl, 10 well ReadyGel, 4-15% Tris-Hcl, 50ul ea ea ea EA EA EA EA $ $ $ $ $ $ $ 9.73 9.73 9.73 8.14 8.14 8.14 8.14 RESTRICTION ENZYMES ...................................................................................................... RE Mfgr. NEB NEB NEB NEB NEB LTI NEB BMB NEB PRO NEB NEB BMB NEB NEB NEB AMR BMB LTI NEB NEB PRO NEB BMB NEB PRO BMB LTI NEB NEB PRO BMB NEB NEB NEB BMB NEB PRO NEB NEB BMB NEB PRO Mfgr No R0117S R0161S R0520S R0541S R0552S 45200-029 R0514S 899208 R0114S R6361 R0558S R0526S 814245 R0153S R0174S R0174L 27-0868-03 567604 15201-023 R0136S R0136L R6021 R0136T 693952 R0160S R6071 348767 15213-010 R0144S R0144M R6081 1292307 R0540S R0517S R0199S 404233 R0162S R6641 R0113S R0524S 404217 R0197S R6551 Code 6101 6104 6107 6312 6410 6108 6414 6346 6111 6396 6288 6323 6405 6116 6385 6416 6120 6117 6118 6119 6297 6121 6124 6126 6128 6398 6131 6281 6298 6408 6133 6407 6299 6387 6381 6348 6137 6400 6384 6388 6138 6139 6289 Item Description Aat II Acc I Afl II Afl III Age I Alu I AlwN I Apa I Apa I Apa I Asc I Ase I Asp 718 Ava II Avr II Avr II BamH I BamH I BamH I BamH I BamH I BamH I BamH I HC Bcl I Bcl I Bgl I Bgl II Bgl II Bgl II Bgl II Bgl II Bsm I BspE I BspH I BssH II BstE II BstE II BstE II BstX I Bsu36 I Cla I Cla I Cla I Unit 500u 500u 2000u 250u 250u 500u 500u 5000u 2000u 5000u 500u 2000u 1000u 2000u 100u 500u 2000u 2500u 2000u 10000u 50000u 2500u 10000u 500u 3000u 1000u 500u 400u 2000u 10000u 500u 200u 1000u 500u 200u 500u 2000u 2000u 1000u 500u 500u 1000u 500u CC Price $ 36.77 $ 44.12 $ 36.77 $ 40.44 $ 40.44 $ 36.70 $ 40.44 $ 47.06 $ 44.12 $ 38.32 $ 40.45 $ 40.45 $ 44.71 $ 36.77 $ 44.12 $ 176.48 $ 26.00 $ 34.51 $ 17.39 $ 36.77 $ 147.07 $ 24.47 $ 36.77 $ 22.75 $ 36.77 $ 30.95 $ 36.87 $ 21.81 $ 36.77 $ 147.07 $ 22.24 $ 81.58 $ 40.44 $ 40.44 $ 40.44 $ 20.40 $ 36.77 $ 38.32 $ 40.44 $ 40.44 $ 42.36 $ 40.44 $ 35.58 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 75 NEB PRO NEB NEB NEB NEB NEB BMB LTI LTI NEB NEB PRO NEB BMB LTI PRO NEB LTI NEB PRO NEB BMB BMB LTI NEB PRO BMB NEB NEB PRO NEB LTI NEB PRO BMB BMB BMB NEB PRO NEB NEB PRO NEB NEB NEB NEB BMB NEB PRO NEB PRO NEB NEB PRO R0175S R6291 R0176S R0129S R0510S R0505S R0521S 703737 15202-013 15202-021 R0101S R0101L R6011 R0101T 667145 15425-010 R6351 R0107S 25212-010 R0139S R6441 R0103S 656313 656321 15207-012 R0104S R6041 798983 R0155S R0105S R6301 R0171S 15232-036 R0142S R6341 742945 822213 862495 R0147S R6711 R0589S R0198S R6381 R0163S R0534S R0190S R0191S 835315 R0193S R6513 R0111S R6801 R0131S R0131L R6501 6142 6401 6140 6389 6390 6143 6300 6144 6145 6336 6147 6301 6149 6152 6156 6157 6290 6159 6162 6325 6358 6166 6168 6354 6169 6303 6172 6174 6176 6180 6361 6182 6339 6185 6363 6184 6394 6417 6190 6364 6324 6123 6365 6413 6193 6305 6411 6199 6201 6203 6204 6367 6205 6308 6206 Dde I Dde I Dpn I Dra I Dra III Eag I (Xma III) EcoN I EcoR I EcoR I EcoR I EcoR I EcoR I EcoR I EcoR I HC EcoR V EcoR V EcoR V Hae II Hha I Hha I Hha I Hinc II Hind III Hind III Hind III Hind III Hind III Hind III HC Hinf I Hpa I Hpa I Hpa II Kpn I Kpn I Kpn I Kpn I HC Mae I Mae II Mbo I Mbo I Mfe I Mlu I Mlu I Mnl I Msc I Nae I Nar I Nco I Nco I Nco I Nde I Nde I Nhe I Nhe I Nhe I 500u 200u 1,000u 2000u 1000u 500u 1000u 5000u 5000u 20000u 10000u 50000u 5000u 10000u 2000u 2000u 2000u 2000u 1500u 2000u 1000u 1000u 5000u 10000u 5000u 10000u 5000u 10000u 5000u 500u 100u 2000u 5000u 3000u 2500u 2000u 50u ea 500u 200u 500u 1000u 1000u 250u 100u 500u 200u 200u 1000u 200u 4000u 500u 1000u 5000u 250u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 36.77 17.81 40.44 40.44 40.44 40.44 40.44 25.89 15.35 66.32 36.77 147.07 17.80 36.77 36.08 28.10 31.14 36.77 49.45 36.77 26.53 36.77 25.10 40.01 14.06 36.77 14.09 40.01 36.77 36.77 29.48 36.77 37.25 36.77 33.17 32.95 76.09 224.35 44.12 36.12 44.13 40.36 35.58 40.44 40.44 36.77 36.77 54.91 36.77 48.92 40.44 66.34 40.44 161.78 40.77 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 76 BMB LTI NEB NEB PRO NEB PRO NEB NEB NEB BMB NEB PRO AMR BMB PRO BMB NEB PRO NEB BMB NEB PRO NEB PRO BMB NEB NEB PRO NEB NEB PRO BMB PRO BMB NEB BMB LTI NEB PRO NEB PRO BMB NEB PRO BMB LTI NEB PRO NEB PRO LTI NEB PRO NEB 1014706 15441-025 R0189S R0189L R6431 R0192S R7091 R0127S R0547S R0560S 621625 R0140S R6111 E1075Y 650137 R6321 642690 R0151S R6331 R0167S 669792 R0156S R6061 R0157S R6221 348783 R0138S R0138L R6051 R0569S R0169S R6191 775258 R6211 1288016 R0123S 220566 15228-018 R0141S R6121 R0130S R6791 1008943 R0133L R6591 1026950 15413-016 R0182S R6261 R0132S R6601 15222-011 R0187S R6421 R0500S 6207 6208 6210 6309 6292 6213 6368 6214 6409 6412 6217 6220 6369 6223 6222 6370 6224 6227 6371 6230 6232 6234 6373 6314 6374 6235 6236 6315 6238 6322 6241 6375 6351 6243 6244 6246 6247 6286 6249 6251 6252 6376 6253 6254 6293 6255 6256 6258 6260 6392 6403 6261 6263 6377 6383 Not I Not I Not I Not I Not I Nru I Nru I Nsi I Pac I Pme I Pst I Pst I Pst I Pvu I Pvu I Pvu I Pvu II Pvu II Pvu II Rsa I Sac I (Sst I) Sac I (Sst I) Sac I (Sst I) Sac II Sac II Sal I Sal I Sal I Sal I Sap I Sau 3A I Sau 3A I Sca I Sca I Sfi I Sfi I Sma I Sma I Sma I Sma I SnaB I SnaB I Spe I Spe I Spe I Sph I Sph I Sph I Sph I Ssp I Ssp I Sst I Stu I Stu I Sty I 200u 200u 500u 2500u 200u 1000u 200u 1000u 250u 500u 3000u 10000u 3000u 150u 100u 100u 1000u 5000u 1000u 1000u 1000u 2000u 1000u 2000u 500u 500u 2000u 10000u 2000u 50u 200u 100u 500u 1000u 250u 2000u 1000u 1000u 2000u 1000u 500u 100u 200u 2500u 200u 200u 200u 250u 200u 1000u 500u 2000u 1000u 400u 3000u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 53.34 30.36 44.12 176.48 35.58 36.77 34.65 40.44 40.44 44.12 25.89 40.44 18.53 53.56 44.71 49.70 31.38 36.77 22.85 36.77 36.08 36.77 29.48 36.77 37.07 36.08 36.77 147.07 38.55 40.45 36.77 24.33 25.89 35.58 17.26 40.44 58.83 44.28 36.77 39.07 36.77 29.48 53.34 161.78 45.21 74.52 64.44 40.44 44.94 40.44 49.39 71.60 36.77 29.48 44.12 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 77 PRO BMB AMR BMB LTI LTI NEB NEB PRO NEB NEB PRO BMB BMB NEB NEB R6481 1371517 27-0948-01 674257 15226-012 15226-038 R0145S R0145L R6181 R0146S R0146L R6161 899194 703770 R0180S R0194S 6404 6406 6272 6268 6269 6344 6271 6320 6379 6274 6287 6380 6352 6277 6393 6280 Sty I Swa I Xba I Xba I Xba I Xba I Xba I Xba I Xba I Xho I Xho I Xho I Xho I Xho I HC Xma I Xmn I 2000u 200u 1500u 1000u 2000u 10000u 3000u 15000u 2000u 5000u 25000u 3000u 5000u 2500u 250u 1000u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 35.38 78.44 28.37 23.53 26.26 113.20 44.12 176.48 38.55 44.12 176.00 24.46 52.55 30.59 40.44 36.77 SERA ................................................................................................. SM Mfgr. SCL GEM LTI EQU SFB BTI HYC LTI HYC Mfgr No FBS-500 100125-500 26140-079 SFB30-1491 F2442-500 FBS-500 SH30071.03 16050-122 SH30087 Code 7138 7137 7141 7140 7135 7133 7139 7123 7126 Item Description Fetal Bovine Serum Fetal Bovine Serum, ES Qualified, Lot # A38507V Fetal Bovine Serum, Lot # 1185797 Fetal Bovine Serum, Lot # SFB30-1491 Fetal Bovine Serum, lot #102K8410 Fetal Bovine Serum, lot #FBU2110 Fetal Bovine Serum, Lot# ANG19178 Horse Serum Iron Enriched Calf Serum — Lot AKH12367 Unit 500 ML 500 ML 500ml 500ml 500 ml 500ML 500ml 500 ml 500 ml CC Price $ 186.88 $ 200.64 $ 241.92 $ 198.90 $ 179.47 $ 156.60 $ 226.86 $ 24.42 $ 51.23 TISSUE CULTURE MEDIA AND REAGENTS.........................................................................TC Mfgr. Mfgr No Code Item Description COL CB40005A 1225 2.5S Nerve Growth Factor - Mouse Natural LTI 12055-091 1228 AIM V Medium LTI 12055-083 1229 AIM V Medium LTI 15240-062 1167 Antibiotic-Antimycotic - Pen/Strep/Fung. LTI 15240-096 1168 Antibiotic-Antimycotic - Pen/Strep/Fung. CCS 1102 1102 Azaserine - 100X 0.6mM 0.1mg/ml COL 354060 1220 bFGF - Human Recombinant LTI 13150-016 1105 Cell Dissociation Buffer - Enzyme free COL CB40236 1201 Collagen - Rat Tail COL CB40235 1262 Dispase CCS 1106 1106 DMEM - high glucose 25mM Hepes LTI 11965-084 1107 DMEM - high glucose 4.5 gm/L MED MT10013CV 1273 DMEM - high glucose 4.5 gm/L MED MT10017CV 1274 DMEM - high glucose 4.5 gm/L - no sodium pyruvate LTI 21063-029 1234 DMEM - high glucose w/o phenol red (1x) LTI 11885-084 1109 DMEM - low glucose LTI 10569-010 1195 DMEM H-glu w/ Glutamax LTI 11320-033 1196 DMEM Nutrient Mix F12 Unit CC Price 0.1 mg $ 500 ml $ 1L $ 100 ml $ 20 ml $ 10 ml $ 10 µg $ 100 ml $ 100 mg $ 100 ml $ 500 ml $ 1L $ 500 ml $ 500 ml $ 500 ml $ 500 ml $ 500ML $ 500 ML $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 78 139.85 32.70 62.22 8.20 3.75 12.00 114.00 13.19 127.06 73.74 10.00 5.96 3.98 3.98 24.61 3.82 4.31 15.23 LTI MED MED LTI LTI LTI BMB LTI BWH LTI MED LTI MED BMB COL LTI LTI BMB COL LTI CCS LTI MED MED MED LTI LTI LTI LTI LTI LTI LTI BWH LTI MED CCS LTI CCS LTI MED MED CCS CCS BMB MED CCS CCS COL BMB COL LTI BMB LTI MED COL 10566-016 MT10013CM MT10017CM 11330-032 10459-014 14080-055 1666789 14200-075 BW17-513Q 14040-117 MT21030CM 14190-136 MT21031CM 855731 CB40001 11130-051 21127-022 1051407 CB40008EA 33016-015 1122 15290-018 MT61234RF MT61234RG MT30-234-CR 11811-031 11811-023 10131-035 15750-011 35050-061 11605-094 11550-043 BW12-615F 11765-047 MT10080CV 1134 14175-079 1137 14170-112 MT21020CM MT21021CM 1139 1279 843555 MT30-240-CR 1140 1141 CB40043 1271164 CB40068 31980-030 1011456 12440-053 MT10016CV 354351 1298 1239 1260 1110 4523 1111 1112 1113 1240 1114 1241 1115 1243 1116 1219 1181 1199 1192 1224 1142 1122 1193 1264 1265 1282 1124 1125 1197 1126 1194 1182 1131 1244 1132 1245 1134 1231 1137 1296 1275 1276 1139 1279 1215 1283 1140 1141 1203 1269 1223 1297 1143 1147 1247 1204 DMEM w/GlutaMAX 500 ML DMEM WITH L-GLUT; HI GLUCOSE; W/ NA PYRUVATE 1 L DMEM WITH L-GLUT; HI GLUCOSE; W/O NA PYRUVATE 1 L DMEM/F-12 - 1:1 mix w/ 15mM Hepes 500 ml DMRIE-C 1 ml DPBS (10X) with Ca++ & Mg++ 500 ml DPBS (10X) without Ca++ & Mg++ 4L DPBS (10X) without Ca++ & Mg++ 500 ml DPBS (1X) with Ca++ & Mg++ 1L DPBS (1X) with Ca++ & Mg++ 1L DPBS (1X) with Ca++ & Mg++ 1L DPBS (1X) without Ca++ & Mg++ 1L DPBS (1X) without Ca++ & Mg++ 1L Epidermal Growth Factor 100 ug Epidermal Growth Factor - Culture Grade 0.1 mg Essential Amino Acids - 50X MEM supplemt 100 ml F12 Nutrient Mix, Kaighns Mod 500 ML Fibronectin - Human 1 mg Fibronectin - Human 1 mg Fibronectin, human plasma 5mg Freezing Mix - 95% Calf Serum 5% DMSO 100 ml Fungizone 20ml G418 1g G418 5g G418 Sulfate (50mg/ml) 20 ml Geneticin (G418)® 5g Geneticin (G418)® 1g Geneticin® - (50mg/ml) 20 ml Gentamicin 10 ml Glutamax I, 100X 100ML Grace's/Hink's supp - yeastolate+lactalbumin 500 ml Ham's F-10 500 ml Ham's F-12 500 ml Ham's F-12 1L Ham's F-12 500 ml Hank's BSS (10X) 500 ml Hank's BSS (1X) w/o Ca++ or Mg++ or phenol 1L Hank's BSS 1X - w/20mM hepes/no Ca&Mg 1L HBSS 500 ML HBSS w/ ca and mg 1L HBSS w/o ca and mg 1L Hybridoma Growth Supp - low endotoxin 10X 25 ml Hybridoma Growth Supplement - High Endotoxin 10–50X 25 ml Hygromycin B (50mg/ml) 20 ml Hygromycin B (50mg/ml) 20 ml Hypoxanthine - 100X 10mM 1.36mg/ml 10 ml Hypoxanthine/Thymidine - 100X 10mM/1.6mM 50 ml IL-2 Human Recombinant 10000u IL-2 Mouse Recombinant 10000u/1ml IL-4 - Human Recombinant 5 µg IMDM w/GlutaMAX 500 ML Interleukin 2 - 10000u Human recombinant 1 ml Iscove's Medium 500 ml Iscove's Medium 500 ml ITS Premix - Culture Supplement 5 ml $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 79 4.09 7.96 7.96 13.41 142.05 9.09 80.80 9.09 6.74 6.36 7.96 6.44 7.96 121.59 92.15 6.44 13.41 87.86 65.87 239.20 22.00 6.10 58.11 201.69 63.10 182.17 56.14 63.66 14.30 5.94 16.22 6.54 5.83 8.56 4.24 12.00 13.73 20.00 8.09 7.96 7.96 24.00 35.00 124.72 97.69 5.00 10.00 93.15 97.27 138.27 6.98 109.04 6.44 6.37 80.72 COL CCS CCS CCS COL LTI BWH MED COL FSH COL COL COL LTI LTI BWH LTI MED LTI LTI LTI LTI LTI LTI BMB LTI LTI IGN CCS BWH LTI MED LTI LTI BWH LTI MED LTI LTI MED LTI LTI LTI LTI LTI CCS MED FSH FSH FSH COL COL LTI MED LTI 354352 1148 1149 1191 CB40232EA 25030-081 BW17-605E MT25005CI 354230 CB40234 CB40230 356237 354253 16600-082 11150-059 BW12-611Q 11095-072 MT10010CM 12571-048 12561-049 12561-056 17504-044 21103-049 11140-050 1011367 09400-72PF 31985-070 IG50 0615 1165 BW17-602E 15140-122 MT30002CI 12040-077 12040-093 BW12-702Q 11875-085 MT10040CM 11835-030 22400-089 MT10041CV 72400-047 15544-018 10902-096 10902-088 25080-094 1175 MT25-000-CI BP2477-1 BP2476-1 BP2473-1 CB40054 CB40039 15090-046 MT25052CI 25300-054 1205 1148 1149 1191 1206 1187 1248 1249 1217 1207 1218 1216 1261 1183 1184 1250 1154 1251 1230 1155 1156 1209 1208 1158 1198 1190 1163 1214 1165 1252 1188 1253 1169 1170 1254 1171 1255 1226 1173 1256 1299 1210 1185 1277 1235 1175 1295 1286 1284 1285 1221 1263 1232 1259 1176 ITS+ Premix - Culture Supplement Kanamycin - 100X - 10mg/ml Kanamycin/Neomycin - 100X - 20 K/5 N mg/ml Kennett's HY - 90% DMEM hg/10% NCTC 135 Laminin - Mouse L-Glutamine - 100X - 200mM - 29.23mg/ml L-Glutamine - 200mM L-Glutamine - 200mM Matrigel® Growth Factor Reduced Matrigel® Matrix Matrigel® Phenol Free & Growth Fact Reduced Matrigel® Phenol Red Free Matrisperse® Cell Recovery Solution McCoy's 5A Medium 199 MEM - w/ Earles' salts & L-Glut MEM - w/ Earles' salts & L-glut MEM - w/ Earles' salts & L-Glut MEM alpha - w/ nucleosides MEM alpha - w/o nucleosides MEM alpha - w/o nucleosides Neural Supplement - B27 (50X) Neuralbasal Media Non-essential amino acids - 100X Nutridoma® SP OPI 100X - 15mg O - 5 mg P - 20u I/ml Opti-MEM® - reduced serum hybrid. medium Origen® Hybridoma Cloning Factor - 10-50X PEG(50%+5%DMSO) - hi efficiency/fusion Pen/Strep - 100X -10000u/ml P - 10mg/ml S Pen/Strep - 100X -10000u/ml P - 10mg/ml S Pen/Strep - 100X -10mcg/ml P - 10mg/ml S PFHM II - Protein Free Hybridoma Medium PFHM II - Protein Free Hybridoma Medium RPMI 1640 RPMI 1640 RPMI 1640 RPMI 1640 - no phenol RPMI 1640 - w/ 25mM Hepes RPMI 1640 - w/ 25mM Hepes RPMI w/GlutaMAX S.O.C. Transformation Media - 10x10 ml SF900 II - Serum-free insect medium SF900 II - Serum-free insect medium Sodium Bicarbonate Sodium Pyruvate - 100X 11g/l 100mM Sodium Pyruvate, 100mM solution TE 10X, pH 7.4 TE, 1X Solution, pH 7.4 TE, 1X Solution, pH 8.0 TGF-alpha - Human Recombinant TGF-beta Human Natural Trypsin - 2.5% (10X) Trypsin-EDTA - 0.05% T - .53mM EDTA Trypsin-EDTA - 0.05% T (1X) 20 ml 10 ml 25 ml 1L 1 mg 100 ml 100 ml 100 ml 10 ml 10 ml 10 ml 10 ml 100 ml 500 ml 500 ml 1L 1L 1L 1L 1L 500 ml 10 ml 500 ml 100 ml 10 ml 50 ml 500 ml 50 ml 2 ml 100 ml 100 ml 100 ml 1L 500 ml 1L 1L 1L 500 ml 500 ml 500 ml 500 ML 100 ml 500 ml 1L 100 ml 100 ml 100 ml 1L 1L 1L 50 µg 1 µg 100 ml 100 ml 100 ml $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 80 60.55 5.00 13.00 16.00 97.10 4.73 9.01 4.77 267.89 223.44 285.80 272.18 51.43 4.55 6.86 8.79 6.59 7.96 27.27 12.63 7.48 42.63 25.64 5.40 33.73 24.13 14.29 146.19 15.00 6.15 4.47 4.77 34.96 20.39 7.68 6.17 7.96 4.90 6.14 5.84 6.45 43.41 19.17 41.98 4.04 5.00 4.30 36.76 25.71 25.61 171.26 244.88 14.29 4.78 3.50 LTI LTI CCS LTI CCS 25200-056 15400-054 1178 11120-052 1180 1236 1177 1178 1179 1180 Trypsin-EDTA - 0.25% (1X) Trypsin-EDTA - 0.5% (10X) Versene - 0.02% EDTA in HBSS Vitamin Mixture - 100X vitamin supplement Water Sterile - Tissue Culture Grade 100 ml 100 ml 1L 100 ml 1L $ $ $ $ $ STOCKED PRODUCTS: VENDOR LISTINGS AMERSHAM BIOSCIENCES, INC. ......................................................................................................... AMR _ Technical Services: (800) 526-3593 _ Website: http://www.amershambiosciences.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: All special orders for Amersham products must be received at the Cell Center by 2pm for next day availability. _ Special Conditions: All non-radioactive items in the Amersham catalog may be special ordered, no shipping or dry ice fees. _ Items ordered on Friday will be available on Monday. _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE MR 4484 27-4005-01 50 bp DNA Ladder 200 µg $ 154.36 CH 2340 27-6200-01 Acetyl CoA 10 mg $ 53.56 MS 4296 NAMP100 Amplify™ 1L $ 143.86 IM 3186 27-4577-01 Anti-GST Antibody 0.5ml $ 207.13 IM 3143 NA931 Anti-Mouse Ig - HRP conjugated 1 ml $ 214.02 IM 3144 NA934 Anti-Rabbit Ig - HRP conjugated 1 ml $ 165.42 PC 5343 27-2056-01 ATP 0.1M 25 µmol $ 48.04 RE 6120 27-0868-03 BamH I 2000u $ 26.00 PC 5345 27-2066-01 CTP 0.1M 25 µmol $ 48.04 PC 5347 27-2050-01 dATP 0.1M 25 µmol $ 47.25 PC 5449 27-2050-02 dATP 100mM solution 100 µm $ 146.49 PC 5348 27-2060-01 dCTP 0.1M 25 µmol $ 47.25 PC 5450 27-2060-02 dCTP 100mM solution 100 µm $ 146.49 PC 5352 27-2045-01 ddNTP Set 4x0.5µmol $ 107.11 CH 2212 17-0340-01 Dextran Sulfate 100 g $ 119.29 PC 5349 27-2070-01 dGTP 0.1M 25 µmol $ 47.25 PC 5451 27-2070-02 dGTP 100mM solution 100 µm $ 146.49 PC 5336 27-2094-01 DNA Polymerization Mix 4x10umol $ 83.48 OE 5373 27-0514-01 DNase 1 1000u $ 63.79 PC 5353 27-2035-02 dNTP Set 4x100umol 4x100µmol $ 519.80 PC 4125 27-2035-01 dNTP Set 0.1M 25 µmol $ 159.88 PC 5350 27-2080-01 dTTP 0.1M 25 µmol $ 42.53 PC 5452 27-2080-02 dTTP 100mM solution 100 µm $ 146.49 NR 5415 RPN2132 ECL™ Plus Western Blotting Detection Reagents 1 ea $ 181.69 NR 3117 RPN2108 ECL™ Western Blot Analysis Starter System 1 kit $ 131.75 NR 3118 RPN2109 ECL™ Western Blotting Detection Reagents 1000cm2 $ 103.33 NR 3119 RPN2106 ECL™ Western Blotting Detection Reagents 4000cm2 $ 225.60 NR 3142 RPN2209 ECL™ Western Blotting Detection Reagents 2000cm2 $ 162.74 OE 5287 E70073Y Exonuclease I 2500u $ 81.94 CH 2126 17-1440-03 Ficoll-Paque® Plus solution 6x500ml $ 337.40 CH 2125 17-1440-02 Ficoll-Paque® solution 6x100ml $ 99.23 KT 5314 27-9261-01 First Strand cDNA Synthesis Kit 1 kit $ 207.13 CH 2213 17-0756-01 Glutathione Sepharose 10 ml $ 159.09 KT 5363 27-4590-01 GST Detection Module 50 rxn $ 182.72 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 81 5.48 6.27 19.00 6.84 9.00 KT PC FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM OE CO CO CO CO OE CO PC CH CO KT RE MR MR MB MB KT OE MB OE OE OE OE OE PC RE 5362 5344 2290 2333 2359 2139 2136 2137 2138 2358 2289 2357 2135 2292 2293 2356 2295 2308 4015 2286 2287 2143 5128 4467 4507 4509 4299 5131 4148 5342 2173 4297 2167 6223 4234 4235 4183 4239 4300 5375 5357 5289 5371 5174 5233 5368 5346 6272 27-4570-01 27-2076-01 RPN2020D RPN303D RPN137N RPN203N RPN2222N RPN303N RPN82N RPN87N RPN303E RPN137B RPN203B RPN2020B RPN303B RPN87B RPN2020F RPN1645 RPN1677L RPN2114K RPN3114K RPN1678K 27-0925-01 27-5325-01 27-5330-01 27-5120-01 27-5130-01 E2420Y 17-0855-02 27-2025-01 17-0891-01 27-5335-01 27-9558-01 E1075Y RPN756 RPN755 27-2166-01 RPN1635 27-9240-01 RPN1633 27-0816-01 E70054Y 27-0808-01 27-0736-01 E2230Z 27-0846-01 27-2086-01 27-0948-01 GST Purifications Bulk Module GTP 0.1M Hybond™ ECL™ 20x20cm Hybond™ ECL™ 30cmx3m Hybond™ Nylon 137mm Hybond™ Nylon 20cmx3m Hybond™ Nylon 22x22cm Hybond™ Nylon 30cmx3m Hybond™ Nylon 82mm Hybond™ Nylon 87mm Hybond™-C extra sup nitrocellulose 30cmx3m Hybond™-N+ charged nylon 137mm Hybond™-N+ charged nylon 20cmx3m Hybond™-N+ charged nylon 20x20cm Hybond™-N+ charged nylon 30cmx3m Hybond™-N+ charged nylon 87mm Hybond™-PVDF 20x20cm Hypercassette™ (beige) 35x43cm Hyperfilm MP Enveloped, 8x10in Hyperfilm™ ECL™ 8x10in Hyperfilm™ ECL™ 8x10in Hyperfilm™ MP 8x10in Maloney-MLV Reverse Transcriptase MicroSpin G-25 Columns MicroSpin G-50 Columns MicroSpin S-200 HR Columns MicroSpin S-300 HR Columns Mung Bean Nuclease NICK® Columns NTP Set 0.1M Percoll® ProbeQuant™ G-50 Micro Columns PureTaq RTG PCR Beads Pvu I Rainbow Marker™ (14300-250000) Rainbow Marker™ (2350-46000) Random Primers (N)6 RapidHyb Hybridization™ Buffer Ready-To-Go DNA Labelling Beads (-dCTP) Rediprime™ kit for dCTP RNAguard Ribonuclease Inhibitor RNase H SP6 RNA Polymerase T4 Polynucleotide Kinase FPLCpure Terminal Deoxy Transferase (TdT) Thrombin Protease UTP 0.1M Xba I 5 prps 25 µmol 10 sht 1 roll 50/bx 1 roll 10/bx 1 roll 50/bx 50/bx 1 roll 50/bx 1 roll 10 sht 1 roll 50/bx 10 sht 1 ea 50 sheets 25 sht 75 sht 75 sht 200u 50 col 50 col 50 col 50 col 3000u 50 col 4x25µmol 1L 50 col 0.5ML 150u 250 µl 250 µl 50 Å260u 125 ml 1 kit 30 rxn 5000u 250u 3000u 200u 2500u 500u 25 µmol 1500u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 266.20 48.04 136.93 168.86 182.30 195.74 157.94 214.22 89.05 99.13 236.06 231.86 200.78 195.74 223.46 126.85 178.51 204.16 144.28 79.55 206.34 206.34 55.13 119.71 119.71 119.71 119.71 118.92 219.73 159.88 226.82 119.71 142.55 53.56 101.12 101.12 117.35 65.37 102.38 186.66 55.13 51.19 152.79 73.24 345.75 62.22 48.04 28.37 ROCHE DIAGNOSTICS .............................................................. BMB _ Technical Services: (800) 262-4911 _ Website: http://www.roche-applied-science.com _ All items listed below are available on site at the Cell Center. University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 82 _ Special Ordering: All items in the Roche catalog may be special ordered at a discount, no shipping or dry ice fees. _ Conditions: All special orders for Roche products must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Monday. _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE KT 5519 1483188 1st Strand cDNA Kit 30 rxn $ MB 4267 1204530 ABTS® Buffer 125 ml $ MB 4266 1112422 ABTS® Tablets (50mg) 20 tab $ CH 2102 101893 Acetyl CoA 10 mg $ CH 2104 1685791 Acrylamide 500 g $ OE 5101 1417215 Agarase 100u $ OE 5374 1758250 Alkaline Phosphatase (shrimp) 1000u $ OE 5103 109118 AMV Reverse Transcriptase 1000u $ IM 3195 1583816 Anti-HA (12CA5) 200 µg $ RE 6346 899208 Apa I 5000u $ RE 6405 814245 Asp 718 1000u $ PC 4103 1140965 ATP 0.1M 40 µmol $ RE 6117 567604 BamH I 2500u $ CH 2379 1383221 BCIP 3ml/150mg $ RE 6126 693952 Bcl I 500u $ CH 2303 874515 Bestatin 10 mg $ RE 6131 348767 Bgl II 500u $ PC 3133 1093070 Biotin-16-dUTP 50 nmol $ CH 2114 1685830 Bis-Acrylamide 25 g $ NR 3154 1096176 Blocking Reagent 50 g $ MB 4104 711454 Bovine Serum Albumin (Nuclease free) 20 mg $ RE 6407 1292307 Bsm I 200u $ RE 6348 404233 BstE II 500u $ OE 5108 713023 Calf Intestinal Alkaline Phosphatase 1000u $ NR 3181 1644807 Cell Proliferation Kit (WST-1) ea $ CH 2115 757306 Cesium Chloride 1 kg $ CH 2116 757292 Cesium Chloride 100 g $ NR 3123 1500708 Chemiluminescent Western Blot Kit 1000cm2 $ RE 6138 404217 Cla I 500u $ MB 4233 1213857 Collagenase P 100 mg $ CH 5522 1873580 Complete EDTA Free 20 Tabs EA $ CH 5521 1836153 Complete Mini 25 Tabs EA $ PC 4105 1140922 CTP 0.1M 40 µmol $ PC 4106 1051440 dATP 0.1M 25 µmol $ PC 6044 1934511 DATP PCR Grade ea $ PC 4108 1051458 dCTP 0.1M 25 µmol $ PC 6045 1934520 DCTP PCR Grade ea $ PC 4115 1051466 dGTP 0.1M 25 µmol $ PC 6046 1934538 DGTP PCR Grade ea $ NR 3178 1585614 DIG/Genius High Prime DNA Labeling and Detection Starter Kit II 1 kit $ NR 3175 1277073 Digoxigenin RNA Labeling Mix 20 rxn $ NR 3151 1209256 Digoxigenin-11-UTP 250 nmol $ CH 2122 100032 Dithiothreitol (DTT) 5g $ MR 4480 1418009 DNA Marker IV (74bp - 19329bp) 50 µg $ MR 4231 1449460 DNA Marker IX (72-1353bp) 50 µg $ MR 4229 821705 DNA Marker V (8-587bp) 50 µg $ MR 4230 1062590 DNA Marker VI (154-2176bp) 50 µg $ MR 4545 1336045 DNA Marker VIII (19bp - 1114bp) 50 µg $ MR 4232 1498037 DNA Marker X (75-12216bp) 100 µg $ MR 4454 1721933 DNA Marker XIV (100 - 1500 bp) 100 bp ladder 50 µg $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 83 236.11 23.53 40.79 52.56 95.70 175.71 69.03 200.81 202.38 47.06 44.71 63.54 34.51 37.65 22.75 99.62 36.87 231.41 26.67 79.23 49.42 81.58 20.40 47.07 278.47 261.21 49.42 76.87 42.36 50.20 225.13 96.48 63.54 47.07 47.07 47.07 47.07 47.07 47.07 192.18 92.56 123.94 54.91 83.15 105.11 83.93 87.86 100.41 87.86 74.52 OE OE PC MB TC PC PC RE RE TC PC PC PC PC PC PC MB PC PC TC CH MB MB PC NR CH MB CH KT KT KT CO CO RE RE RE TC TC NR TC CH OE RE PC PC CH NR NR RE RE CH RE RE TC PC 5320 5114 6043 4126 1112 4127 6047 6144 6156 1116 5325 6011 6018 5274 6003 6012 5440 5442 5443 1192 2127 5421 5422 5444 3130 2131 4131 2134 5281 5282 5520 4512 4513 6168 6354 6174 1215 1269 3173 1143 2145 5125 6184 5523 5524 2146 6041 6042 6394 6417 2378 6199 6207 1198 5327 104159 776785 1969064 1202375 1666789 1051482 1934546 703737 667145 855731 1732641 1732650 1759078 1681834 1759060 1681842 3045595 2032902 2032929 1051407 1814320 1815091 1814443 2140306 1175025 100149 901393 1685929 1585584 1585592 1796828 1754777 1754785 656313 656321 798983 843555 1271164 1684795 1011456 724815 1008404 742945 2239264 3003230 1017128 2015200 2015196 822213 862495 1383213 835315 1014706 1011367 1696513 DNase I - Grade II DNase I - RNase Free dNTP's PCR Grade DOTAP (Liposomal Transfection Reagent) DPBS (10X) without Ca++ & Mg++ dTTP 0.1M DTTP PCR Grade EcoR I EcoR V Epidermal Growth Factor Expand™ High Fidelity PCR System Expand™ High Fidelity PCR System Expand™ High Fidelity PCR System Expand™ Long Template PCR System Expand™ Long Template PCR System Expand™ Long Template PCR System ExtremeGENE (Transfection Reagent) FastStart Taq DNA Polymerase FastStart Taq DNA Polymerase Fibronectin - Human Formamide FuGENE 6 (Nonliposomal Transfection Reagent) FuGENE 6 (Nonliposomal Transfection Reagent) GC Rich Polymerase GENIUS™ 4 RNA Labeling Kit Glycine Glycogen Guanidine Thiocyanate High Prime DNA Labeling Kit High Prime DNA Labeling mix for dCTP High Pure PCR Template Prep High Pure Plasmid Isolation Kit High Pure Plasmid Isolation Kit Hind III Hind III Hind III HC Hygromycin B (50mg/ml) IL-2 Mouse Recombinant In Situ Cell Death Detection Kit - Fluorescein Interleukin 2 - 10000u Human recombinant IPTG Klenow - Labeling grade Kpn I HC LC FastStart DNA Mstr Sybrgreen LC FastStart DNA Mstr Sybrgreen Leupeptin Lumi Light Lumi Light + Mae I Mae II NBT (Nitro Blue Tetra) Nco I Not I Nutridoma® SP PCR Cleanup Kit 100 mg 10000u ea 2 ml 4L 25 µmol ea 5000u 2000u 100 ug 100u 500u 10x250u 100u 10x250u 500u .4 ml 100u 500u 1 mg 500 ml .4 ml 1 ml 50 rxn 2x10rxn 1 kg 20 mg 500 g 50 rxn 50 rxn EA 50 col 250 col 5000u 10000u 10000u 20 ml 10000u/1ml 50 rxn 1 ml 1g 100u 2000u EA EA 25 mg 1 kit 1 kit 50u ea 3 ml 200u 200u 10 ml 100 rxn $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 84 36.08 109.04 165.51 242.39 80.80 47.07 47.07 25.89 36.08 121.59 69.03 324.75 1,455.11 73.74 1,546.10 342.01 84.72 59.62 283.18 87.86 36.08 125.51 250.23 146.69 311.42 40.79 77.66 135.71 207.09 188.26 131.78 47.85 206.30 25.10 40.01 40.01 124.72 97.27 266.70 109.04 32.16 47.85 32.95 828.35 207.09 105.11 142.77 247.09 76.09 224.35 36.87 54.91 53.34 33.73 111.39 PC PC CH CH CH CH MB CH MB CH CH CH CH RE RE RE CO CO CO CO CO KT MB OE OE OE RE RE RE RE NR RE CH RE RE CH CH RE OE OE OE OE PC PC PC PC CH PC PC PC PC PC CH CH CH 4197 4289 2377 2172 2237 2174 4152 2345 5525 5136 2444 5518 5526 6217 6222 6224 4165 4166 4167 4326 4168 5139 4170 5143 5386 5388 6232 6235 6351 6244 3132 6247 2185 6253 6255 2186 2189 6406 5163 5167 5259 5171 5185 5198 6002 5199 2196 5461 5462 5463 5885 5988 2198 2199 2203 1581295 1636138 1429868 1359053 100995 1814303 108812 1697498 3335399 3115879 1373200 1000144 3115828 621625 650137 642690 1273922 1273990 1273965 1273973 1274015 1004760 1034731 1119915 109169 109193 669792 348783 775258 1288016 1093274 220566 1667262 1008943 1026950 1666681 100168 1371517 481220 1004786 1635379 709557 1146165 NA 1435094 1578553 100139 3186172 3186202 3186199 1855476 1888382 604205 604203 789704 PCR Nucleotide Mix PCR Optimization Kit (for optimal pH & Mg) Pefabloc® SC Pepstatin (was #600160) Phenol Saturated (Tris) Phenol (was 100300) Poly d(I-C) Protease Inhibitor Cocktail Tablets (Complete) Protector RNase Inhibitor Proteinase K Proteinase K Proteinase K Proteinase K Solution (20 mg/ml) Pst I Pvu I Pvu II Quick Spin™ G25 DNA Quick Spin™ G25 RNA Quick Spin™ G50 DNA Quick Spin™ G50 DNA Quick Spin™ G50 RNA Random Primed DNA Labeling Kit Random Primers (N)6 RNase - DNase free Rnase A RNase T1 Sac I (Sst I) Sal I Sca I Sfi I Sheep Anti-digoxigenin-AP Sma I Sodium Dodecyl Sulfate Spe I Sph I SSC 20X Sucrose Swa I T4 DNA Ligase T4 DNA Polymerase T4 DNA Rapid Ligation Kit (5 minutes) T4 Polynucleotide Kinase Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq PCR Core Kit TEMED Tgo DNA Polymerase Tgo DNA Polymerase Tgo DNA Polymerase TITAN™ One-Tube RT-PCR System TITAN™ One-Tube RT-PCR System Tris Tris Triton® X-100 100 rxn 1 kit 100 mg 10 mg 100 ml 500 g 10 Å260u 20 tabs EA 100 mg 25 ml 500 mg 5 ml 3000u 100u 1000u 20 col 20 col 20 col 50 col 20 col 50 rxn 50 Å260u 500 µg 100 mg 100000u 1000u 500u 500u 250u 150u 1000u 100 g 200u 200u 4L 1 kg 200u 100u 100u 40 rxn 200u 100u 250u 5000u 100 rxn 50 g 50 rxns 100 rxns 250 rxns 100 rxn 25 rxn 1 kg 500 g 100 ml $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 85 49.42 123.15 90.99 47.07 48.63 92.56 66.68 225.13 64.32 59.61 313.77 61.97 90.99 25.89 44.71 31.38 71.38 103.54 71.38 133.35 103.54 209.44 111.39 57.26 57.26 33.73 36.08 36.08 25.89 17.26 127.86 58.83 29.81 53.34 74.52 80.80 32.16 78.44 40.79 73.74 104.33 160.81 53.34 65.00 1,255.08 185.91 32.16 93.35 156.89 327.11 336.52 105.90 57.26 36.08 25.10 OE NR NR RE CH CH RE RE 5387 3180 3179 6268 2205 2206 6352 6277 109541 1767305 1767291 674257 745740 651745 899194 703770 tRNA TUNEL Enzyme (optimized terminal transferase) TUNEL Label (FITC dUTP + dNTPs) Xba I X-gal X-gal Xho I Xho I HC 100 mg 1 ea 1 ea 1000u 1g 250 mg 5000u 2500u $ $ $ $ $ $ $ $ BIOWHITTAKER MOLECULAR APPLICATIONS ............................................................……………………….. BMA _ Technical Services: (800) 654-4452 _ Website: http://www.biowhittaker.com; http://www.bmaproducts.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: No. _ Delivery: 3rd Business Day PC CODE MFGR. NO. DESCRIPTION TC 1240 BW17-513Q DPBS (1X) with Ca++ & Mg++ TC 1244 BW12-615F Ham's F-12 TC 1248 BW17-605E L-Glutamine - 200mM TC 1250 BW12-611Q MEM - w/ Earles' salts & L-Glut TC 1252 BW17-602E Pen/Strep - 100X -10000u/ml P - 10mg/ml S TC 1254 BW12-702Q RPMI 1640 SIZE 1L 500 ml 100 ml 1L 100 ml 1L CC PRICE $ 6.74 $ 5.83 $ 9.01 $ 8.79 $ 6.15 $ 7.68 CAMBREX BIOSCEINCES …………………………………………………………………….FMC _ Technical Services: 800-341-1574 _ All items listed below are available on site at the Cell Center. _ Special Orders: All items in the Cambrex catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Wednesday. _ Delivery: 3rd Business Day. PC CH CH CH CH CH CH CODE 2234 2263 2321 2233 2236 2235 MFGR. NO. 50100 50101 50090 50080 50004 50000 DESCRIPTION Agarose (LMP) SeaPlaque® Agarose (LMP) SeaPlaque® Agarose NuSieve® 3:1 Agarose NuSieve® GTG® Agarose SeaKem® LE Agarose SeaKem® LE SIZE 125 g 25 g 125 g 125 g 500 g 125 g CC PRICE $ 289.24 $ 105.93 $ 223.98 $ 278.01 $ 285.54 $ 111.25 CELL CENTER SERVICES ..................................................................................................... CC _ Technical Services: (215) 898-2795 _ Website: http://www.med.upenn.edu/ccsf/ _ All items listed below are available on site at the Cell Center. _ Special Ordering: Please contact the Cell Center Service Facilty at 898-2795 for any special requests. _ All items have been produced by the Cell Center, except for Xgal which is purchased from Diagnostic Chemicals, Ltd. and is packaged under nitrogen. It is of high purity and quality and has been tested in screening assays and histological staining procedures. PC CODE DESCRIPTION SIZE CC PRICE University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 86 92.56 92.56 69.03 23.53 147.47 86.29 52.55 30.59 CH 2111 Ampicillin - 25mg/ml 10 ml $ 10.00 TC 1102 Azaserine - 100X 0.6mM 0.1mg/ml 10 ml $ 12.00 CH 2117 Chloramphenicol 34 mg/ml 10 ml $ 8.00 TC 1106 DMEM - high glucose 25mM Hepes 500 ml $ 10.00 TC 1122 Freezing Mix - 95% Calf Serum 5% DMSO 100 ml $ 22.00 TC 1134 Hank's BSS (10X) 500 ml $ 12.00 TC 1137 Hank's BSS 1X - w/20mM hepes/no Ca&Mg 1L $ 20.00 TC 1139 Hybridoma Growth Supp - low endotoxin 10X 25 ml $ 24.00 TC 1279 Hybridoma Growth Supplement - High Endotoxin 10–50X 25 ml $ 35.00 TC 1140 Hypoxanthine - 100X 10mM 1.36mg/ml 10 ml $ 5.00 TC 1141 Hypoxanthine/Thymidine - 100X 10mM/1.6mM 50 ml $ 10.00 TC 1148 Kanamycin - 100X - 10mg/ml 10 ml $ 5.00 TC 1149 Kanamycin/Neomycin - 100X - 20 K/5 N mg/ml 25 ml $ 13.00 TC 1191 Kennett's HY - 90% DMEM hg/10% NCTC 135 1L $ 16.00 CH 6005 LB + Ampicillin (50ug/ml) Agar Plates 10 pl $ 10.00 CH 6006 LB + Kanamycin (50ug/ml) Agar Plates 10 pl $ 10.00 CH 6007 LB + Tetracycline (20ug/ml) Agar Plates 10 pl $ 10.00 CH 6004 LB Agar Plates 10 pl $ 10.00 IM 3207 myc 1 - 9E10 antibody 0.5 mg $ 20.00 TC 1165 PEG(50%+5%DMSO) - hi efficiency/fusion 2 ml $ 15.00 MB 4463 Primer gt10 forward 5 µg $ 20.00 MB 4464 Primer gt10 reverse 5 µg $ 20.00 MB 4465 Primer gt11 forward 5 µg $ 20.00 MB 4466 Primer gt11 reverse 5 µg $ 20.00 MB 4190 Primer M13/pUC Forward 23mer 5 µg $ 20.00 MB 4191 Primer M13/pUC Reverse 23mer 5 µg $ 20.00 MB 4194 Primer Sp6 20mer 5 µg $ 20.00 MB 4192 Primer T3 20mer 5 µg $ 20.00 MB 4193 Primer T7 20mer 5 µg $ 20.00 MB 4524 Primer T7 Term 5 µg $ 20.00 TC 1175 Sodium Pyruvate - 100X 11g/l 100mM 100 ml $ 5.00 CH 2197 Tetracycline 12.5mg/ml 10 ml $ 8.00 TC 1178 Versene - 0.02% EDTA in HBSS 1L $ 19.00 TC 1180 Water Sterile - Tissue Culture Grade 1L $ 9.00 CH 2207 X-gal 1g $ 65.00 CLONTECH LABORATORIES ............................................................................................. CLN _ Technical Services: 800-662-CLON _ Website: http://www.clontech.com/ _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: All items in the Clontech catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders for CLN products must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Tuesday. _ Delivery: Next Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE PC 5985 639102 Advantage cDNA PCR Kit 1 ea $ 139.34 PC 5987 639105 Advantage cDNA Plolymerase Mix 1 ea $ 254.83 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 87 PC KT CO CO CH CH CH CH MB CH CH MB MB MB MB 6051 6008 4533 4536 5992 5989 5981 5991 6026 5905 5903 6027 6028 6030 6029 639505 631312 636055 636060 630415 630414 630413 630416 636831 630412 630411 635501 635506 630440 630439 Advantage RT for PCR Cal Phos Mammalian Transfection Kit CHROMA SPIN-10 Columns CHROMA SPIN-100 Columns DO Supplement-His DO Supplement-Leu DO Supplement-Trp DO Supplement-Ura Express Hyb Minimal SD Agar Base Minimal SD Base Talon Metal Affinity Resin Talon Superflow Metal Affinity Resin Yeastmaker Carrier DNA Yeastmaker Yeast Transformation System ea sm box 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 250 ml 1 ea 1 ea 10 ml 25 ml ea kit $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 241.50 143.63 112.10 112.12 34.29 37.07 34.29 34.29 148.63 139.00 92.66 69.64 194.59 199.23 203.86 SIZE 0.1 mg 10 µg 100 mg 100 ml 0.1 mg 1 mg 10000u 5 µg 5 ml 20 ml 1 mg 10 ml 10 ml 10 ml 100 ml 50 µg 1 µg 10µg CC PRICE $ 139.85 $ 114.00 $ 127.06 $ 73.74 $ 92.15 $ 65.87 $ 93.15 $ 138.27 $ 80.72 $ 60.55 $ 97.10 $ 267.89 $ 285.80 $ 272.18 $ 51.43 $ 171.26 $ 244.88 $ 225.88 COLLABORATIVE BIOMEDICAL PRODUCTS ................................................................... COL _ Technical Services: (800) 343-2035 _ Website: http://www.bd.com/ _ All items listed below are available on site at the Cell Center. _ Special Orders: No. PC CODE MFGR. NO. DESCRIPTION TC 1225 CB40005A 2.5S Nerve Growth Factor - Mouse Natural TC 1220 354060 bFGF - Human Recombinant TC 1201 CB40236 Collagen - Rat Tail TC 1262 CB40235 Dispase TC 1219 CB40001 Epidermal Growth Factor - Culture Grade TC 1224 CB40008EA Fibronectin - Human TC 1203 CB40043 IL-2 Human Recombinant TC 1223 CB40068 IL-4 - Human Recombinant TC 1204 354351 ITS Premix - Culture Supplement TC 1205 354352 ITS+ Premix - Culture Supplement TC 1206 CB40232EA Laminin - Mouse TC 1217 354230 Matrigel® Growth Factor Reduced TC 1218 CB40230 Matrigel® Phenol Free & Growth Fact Reduced TC 1216 356237 Matrigel® Phenol Red Free TC 1261 354253 Matrisperse® Cell Recovery Solution TC 1221 CB40054 TGF-alpha - Human Recombinant TC 1263 CB40039 TGF-beta Human Natural IM 3208 CB40066 TNF-a DENVILLE SCIENTIFIC (Q-BIOGENE / TKR) ........................................................................................................DEN _ Technical Services: (800) 453-0385 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: Items in the Denville Scientific catalog may be special ordered at a discount, no shipping or dry ice fees. _ Conditions: Items listed in the Supplies and Chemicals sections of the Denville catalog may be special ordered. _ Delivery: 3rd Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE MS 4462 R6651 Chillete personal tube cooler -12 tubes at -20C 1 ea $ 28.60 PC 3307 CB-4420-2 dNTP Set (4x25umol) 100mM EA $ 130.45 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 88 CO 4614 1102-200 GeneClean Turbo 50 prp $ 57.36 CO 4450 1101-200 GeneClean® DNA Purification Spin Kit 50 prp $ 78.53 CO 4569 1101-400 GeneClean® DNA Purification Spin Kit 100 prp $ 139.44 CO 4269 1001-400 GeneClean® II DNA Purification Kit 300 prp $ 123.53 CO 4448 1001-600 GeneClean® III DNA Purification Kit 600 prp $ 180.01 CO 4451 2055-400 Lambda Quick! Phage DNA Isolation Spin Kit 25 prp $ 100.60 CH 2420 3002-031 LB Medium Capsules 1 kg $ 107.64 MS 4621 TKR-1301 Mini-Centrifuge ea $ 250.00 MS 4497 C1201-CL Mini-Centrifuge 6 x 1.5 ml 1 ea $ 185.59 MS 3327 P-1125 P-100 Filter Pipet Tips EA $ 47.30 MS 3329 P-1123 P-1000 Filter Pipet Tips EA $ 47.30 MS 3325 P-1096-FR P2/P10 Eatra Long Filter Pipet Tip EA $ 47.30 MS 3326 P-1121 P-20 Filter Pipet Tips EA $ 47.30 MS 3328 P-1122 P-200 Filter Pipet Tips EA $ 47.30 MS 4456 P3950-10 Pipet-mate Digital Pipette w/ tip ejector 10µl 1 ea $ 158.95 MS 4458 P3950-100 Pipet-mate Digital Pipette w/ tip ejector 100µl 1 ea $ 152.01 MS 4460 P3950-1000 Pipet-mate Digital Pipette w/ tip ejector 1000µl 1 ea $ 152.01 MS 4457 P3950-20 Pipet-mate Digital Pipette w/ tip ejector 20µl 1 ea $ 152.01 MS 4459 P3950-200 Pipet-mate Digital Pipette w/ tip ejector 200µl 1 ea $ 152.01 MS 4461 P3950-5000 Pipet-mate Digital Pipette w/ tip ejector 5000µl 1 ea $ 159.29 DIFCO LABORATORIES (FISHER)........................................................................................DIF _ Technical Services: (800) 343-2035 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION CH 5889 DF0145170 Agar - Granulated CH 5890 DF0230173 Casamino Acids CH 5891 DF0445174 LB Agar - Miller CH 5893 DF0402170 LB Broth - Lennox CH 5892 DF0446173 LB Broth - Miller CH 5886 DF0118170 Peptone CH 5887 DF0123173 Tryptone Peptone CH 5888 DF0127179 Yeast Extract SIZE 500 g 500 g 500 g 500 g 500 g 500 g 500 g 500 g CC PRICE $ 59.09 $ 51.84 $ 44.71 $ 20.43 $ 20.43 $ 41.30 $ 36.39 $ 51.68 ENZO LIFE SCIENCES, INC. ……………………………………………………………………….ENZ _ Technical Services (800) 221-7705 _ All items below are available on site at the Cell Center. _ Special Ordering: No. _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION KT 4630 42655-10 BioArray High Yield RNA Transcript Labeling Kit KT 4631 42655-20 BioArray High Yield RNA Transcript Labeling Kit SIZE 10 rxn 20 rxn CC PRICE $ 705.00 $ 1,380.75 EPICENTRE TECHNOLOGIES University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 89 ...............................................................................................EPI _ Technical Services: (800) 284-8474 _ Web Site: http://www.epicentre.com _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION OE 5322 SL910K SP6 RNA Polymerase (25u/µl) SIZE 10000u CC PRICE $ 102.02 FISHER SCIENTIFIC ............................................................................................................. FSH _ Technical Services: (800) 533-8355 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION CH 2224 BP170500 Acrylamide CH 4308 BP1423500 Agar - Granulated MS 6023 121311 Aluminum Foil (18” x 50ft) CH 4337 BP179100 Ammonium Persulfate MS 4222 212363 ART® 10 aerosol resistant pipet tip (racked) MS 4228 212362A ART® 1000E aerosol resistant pipet tip (racked) MS 4226 212368 ART® 100E aerosol resistant pipet tip (racked) MS 4227 212361 ART® 200 aerosol resistant pipet tip (racked) MS 4224 212365 ART® 20E aerosol resistant pipet tip (racked) MS 4223 212364 ART® 20P aerosol resistant pipet tip (racked) CH 2405 BP1610100 BCIP CH 4338 BP17125 Bis-Acrylamide CH 2406 BP168500 Boric Acid CH 4309 BP1424500 Casamino Acids CH 2226 BP210500 Cesium Chloride FM 2118 57144 Chromatography Paper 3MM 46X57cm CH 4339 BP339500 Citric Acid CH 4440 BP1585100 Dextran Sulfate CH 2239 BP1725 Dithiothreitol (DTT) CH 2407 BP2311 DMSO CH 2408 BP120500 EDTA Disodium Salt Dihydrate CH 2227 BP227500 Formamide CH 2230 BP3811 Glycine CH 4442 BP221250 Guanidine Isothiocyanate CH 4441 BP310100 HEPES PC 5514 E954140156 HotMaster Taq w/ buffer PC 5515 E954140164 HotMaster Taq w/ buffer PC 5516 E954140172 HotMaster Taq w/ buffer CH 2238 BP16201 IPTG CH 5299 BP162010 IPTG CH 4310 BP1425500 LB Agar - Miller CH 4311 BP1427500 LB Broth - Lennox CH 4312 BP1426500 LB Broth - Miller CH 2410 BP214500 Magnesium Chloride PC 5513 E0032002250 MasterMix PC 5510 E0032002404 MasterTaq Kit PC 5511 E0032002501 MasterTaq Kit PC 5512 E0032002552 MasterTaq Kit SIZE 500 g 500 g ea 100 g 960/pk 800/pk 960/pk 960/pk 1000/pk 960/pk 100 mg 25 g 500 g 500 g 500 g 100/pk 500 g 100 g 5g 1L 500 g 500 ml 1 kg 250 g 100 g 100u 250u 1000u 1g 10 g 500 g 500 g 500 g 500 g 152u 100u 250u 500u CC PRICE $ 74.59 $ 68.93 $ 8.53 $ 15.24 $ 57.00 $ 57.00 $ 57.00 $ 57.00 $ 56.98 $ 57.00 $ 18.65 $ 19.44 $ 17.59 $ 46.88 $ 131.63 $ 85.24 $ 15.72 $ 87.37 $ 42.03 $ 30.36 $ 30.64 $ 30.90 $ 29.35 $ 58.61 $ 32.36 $ 31.95 $ 74.55 $ 266.25 $ 27.17 $ 202.45 $ 54.23 $ 26.88 $ 26.94 $ 22.37 $ 39.94 $ 22.37 $ 55.91 $ 106.50 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 90 TC CH CH CH CH CH CH CH CH CH MB CH CH PC PC TC TC TC CH CH CH CH CH CH CH MS CH 1207 4313 2231 5300 5301 5302 5303 2411 5211 5304 4201 2232 4443 5309 5311 1286 1284 1285 4444 5305 2412 2413 4314 2240 2409 6024 4315 CB40234 BP1420500 BP226500 BP1750I100 BP1750I400 BP1752I100 BP1752I400 BP366500 BP1700100 BP1700500 2123621 BP166500 BP2201 FB600010 FB600045 BP2477-1 BP2476-1 BP2473-1 BP1521 BP13381 BP152500 BP13071 BP1421500 BP169500 BP5611 15610 BP1422500 Matrigel® Matrix Peptone Phenol Phenol Saturated pH 6.6 Phenol Saturated pH 6.6 Phenol/Chloroform/Isoamyl Alcohol Phenol/Chloroform/Isoamyl Alcohol Potassium Chloride Proteinase K Proteinase K RNase AWAY Sodium Dodecyl Sulfate (SDS) Sucrose Taq DNA Polymerase w/ buffer A Taq DNA Polymerase w/ buffer B TE 10X, pH 7.4 TE, 1X Solution, pH 7.4 TE, 1X Solution, pH 8.0 Tris Base TRIS EDTA Solution 100X Tris-Base Tris-Glycine (10X) Tryptone Urea Water(Sterile Nuclease Free) Wrapping Film (18in x 1000ft) Yeast Extract 10 ml 500 g 500 g 100 ml 400 ml 100 ml 400 ml 500 g 100 mg 500 mg 8.4 oz 500 g 1 kg 100u 100u 1L 1L 1L 1 kg 1L 500 g 1L 500 g 500 g 1L ea 500 g $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 223.44 46.06 63.92 33.30 71.96 30.24 72.45 15.18 61.80 285.55 17.05 61.80 20.57 149.17 149.17 36.76 25.71 25.61 51.75 55.41 32.76 31.44 37.83 21.21 17.59 18.66 54.35 FUJI FILM ................................................................................................................FUJ _ Technical Services: (800) 533-8355 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION FM 2325 04-441-118 Fuji RX Film NIF 14x17in FM 2324 04-441-115 Fuji RX Film NIF 8x10in SIZE 100 sht 100 sht CC PRICE $ 305.28 $ 111.69 GEMINI BIOPRODUCTS......................................................................................GEM _ Technical Services: (800) 543-6464 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Business Day PC CODE MFGR. NO. DESCRIPTION SM 7137 100125-500 Fetal Bovine Serum, ES Qualified, Lot # A38507V SIZE 500 ML CC PRICE $ 200.64 HYCLONE LABORATORIES ......................................................................………..........................HYC _ Technical Services: (800) 492-5663 _ All items listed below are available on site at the Cell Center. _ Delivery: Overnight University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 91 PC SM SM CODE MFGR. NO. 7139 SH30071.03 7126 SH30087 DESCRIPTION Fetal Bovine Serum, Lot# ANG19178 Iron Enriched Calf Serum — Lot AKH12367 SIZE 500ml 500 ml CC PRICE $ 226.86 $ 51.23 IGEN INTERNATIONAL (FISHER) ........................................................................................ IGN _ Technical Services: (800) 533-8355 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION TC 1214 IG50 0615 Origen® Hybridoma Cloning Factor - 10-50X SIZE 50 ml CC PRICE $ 146.19 INVITROGEN (GIBCO LIFE TECHNOLOGIES) ................................................................................. INV _ Technical Services: (800) 955-6288 _ Web Site: http://www.invitrogen.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: All items in the Invitrogen / Life Technologies catalog may be special ordered at a discount, no shipping or dry ice fees. Freezer items ordered on Friday will be available on Tuesday. _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE MR 4123 15615-016 1 kb DNA Ladder 250 µg $ 93.19 MR 4221 15615-024 1 kb DNA Ladder 1000 µg $ 309.87 MR 4302 10381-010 1 kb DNA Ladder - RTS - ready load 100 app $ 53.13 MR 4625 10787-018 1 kb Plus DNA Ladder 250 µg $ 84.88 PC 4010 18427-088 10 mM dNTP Mix 1 ML $ 216.22 MR 4101 15628-019 100 bp DNA Ladder 50 µg $ 41.70 MR 4304 10380-012 100 bp Ladder - RTS - ready load 100 app $ 64.05 MR 4196 10068-013 100 bp-2kb DNA Mass Ladder 200 µl $ 64.41 PC 5468 18427013 10mM dNTP Mix 100 µl $ 28.61 MR 4102 15613-011 123 Base Pair Ladder 100 µg $ 67.10 MR 4498 10597-011 25 bp DNA Ladder 50 µg $ 71.58 CH 5453 10812-014 2nd Strand Buffer 0.5 ml $ 53.89 PG 6068 EA0375 3-8% NuPAGE® Tris-Acetate Gels 1.0 mm 10 well ea $ 9.73 PG 6069 EA03752 3-8% NuPAGE® Tris-Acetate Gels 1.0 mm 12 well ea $ 9.73 PG 6070 EA0378 3-8% NuPAGE® Tris-Acetate Gels 1.5 mm 10 well ea $ 9.73 MR 4499 10416-014 50 bp DNA Ladder 50 µg $ 71.58 PG 6071 EA0355 7% NuPAGE® Tris-Acetate Gels 1.0 mm 10 well ea $ 9.73 PG 6072 EA0358 7% NuPAGE® Tris-Acetate Gels 1.5 mm 10 well ea $ 9.73 CH 4185 30391-023 Agar - Bacterial Select 500 g $ 64.27 CH 2105 15510-019 Agarose 100 g $ 73.39 CH 2106 15510-027 Agarose 500 g $ 256.80 CH 1289 10975-035 Agarose 1000 100G $ 181.90 TC 1228 12055-091 AIM V Medium 500 ml $ 32.70 TC 1229 12055-083 AIM V Medium 1L $ 62.22 RE 6108 45200-029 Alu I 500u $ 36.70 TC 1167 15240-062 Antibiotic-Antimycotic - Pen/Strep/Fung. 100 ml $ 8.20 TC 1168 15240-096 Antibiotic-Antimycotic - Pen/Strep/Fung. 20 ml $ 3.75 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 92 IM 4636 R96025 Anti-V5 Antibody 50 ul $ 148.21 RE 6118 15201-023 BamH I 2000u $ 17.39 MR 4619 10748-010 BENCHMARK™ Prestained Protein Ladder 2 x 250u $ 78.71 RE 6281 15213-010 Bgl II 400u $ 21.81 CH 4635 R21001 Blasticidin 50 MG $ 140.80 MB 4618 15561-020 Bovine Serum Albumin, Acetylated 150 mg $ 98.67 TC 1105 13150-016 Cell Dissociation Buffer - Enzyme free 100 ml $ 13.19 CE 4118 18263-012 DH5alpha™ Library Efficiency Comp Cells 1 ml $ 94.88 CE 4119 18265-017 DH5alpha™ Subcloning Efficiency Comp Cells 2 ml $ 43.69 TC 1107 11965-084 DMEM - high glucose 4.5 gm/L 1L $ 5.96 TC 1234 21063-029 DMEM - high glucose w/o phenol red (1x) 500 ml $ 24.61 TC 1109 11885-084 DMEM - low glucose 500 ml $ 3.82 TC 1195 10569-010 DMEM H-glu w/ Glutamax 500ML $ 4.31 TC 1196 11320-033 DMEM Nutrient Mix F12 500 ML $ 15.23 TC 1298 10566-016 DMEM w/GlutaMAX 500 ML $ 4.09 TC 1110 11330-032 DMEM/F-12 - 1:1 mix w/ 15mM Hepes 500 ml $ 13.41 TC 4523 10459-014 DMRIE-C 1 ml $ 142.05 OE 5183 18010-025 DNA Polymerase I (E Coli) 1000U $ 113.64 OE 5113 18047-019 DNase I 20000u $ 184.22 OE 5225 18068-015 DNase I - Amplification Grade 100u $ 56.95 TC 1111 14080-055 DPBS (10X) with Ca++ & Mg++ 500 ml $ 9.09 TC 1113 14200-075 DPBS (10X) without Ca++ & Mg++ 500 ml $ 9.09 TC 1114 14040-117 DPBS (1X) with Ca++ & Mg++ 1L $ 6.36 TC 1115 14190-136 DPBS (1X) without Ca++ & Mg++ 1L $ 6.44 OE 5454 18052-019 E. Coli DNA Ligase 100u $ 23.20 RE 6145 15202-013 EcoR I 5000u $ 15.35 RE 6336 15202-021 EcoR I 20000u $ 66.32 RE 6157 15425-010 EcoR V 2000u $ 28.10 CH 2283 15575-020 EDTA 0.5M (4 x 100ml) 400 ml $ 23.91 CE 5423 18290-015 ElectroMAX DH10B Efficiency Comp Cells 5x.1ml $ 156.66 CE 5426 11319-019 ElectroMAX DH5alpha-E Efficiency Comp Cells 5x.1ml $ 154.63 MS 4519 P410-50 Electroporation Cuvettes 0.1cm $ 115.26 MS 4520 P450-50 Electroporation Cuvettes 0.2cm $ 115.26 MS 4521 P460-50 Electroporation Cuvettes 0.4cm $ 115.26 OE 5475 10480-010 Elongase Enzyme Mix 100 rxn $ 100.62 TC 1181 11130-051 Essential Amino Acids - 50X MEM supplemt 100 ml $ 6.44 CH 2277 15585-011 Ethidium Bromide (ETBR) 10mg/ml 10 ml $ 28.47 TC 1199 21127-022 F12 Nutrient Mix, Kaighns Mod 500 ML $ 13.41 CO 5221 K1593-02 FastTrack® 2.0 Kit 6 rxn $ 322.10 SM 7141 26140-079 Fetal Bovine Serum, Lot # 1185797 500ml $ 241.92 TC 1142 33016-015 Fibronectin, human plasma 5mg $ 239.20 TC 1193 15290-018 Fungizone 20ml $ 6.10 TC 1124 11811-031 Geneticin (G418)® 5g $ 182.17 TC 1125 11811-023 Geneticin (G418)® 1g $ 56.14 TC 1197 10131-035 Geneticin® - (50mg/ml) 20 ml $ 63.66 TC 1126 15750-011 Gentamicin 10 ml $ 14.30 TC 1194 35050-061 Glutamax I, 100X 100ML $ 5.94 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 93 CH 2130 15514-011 Glycerol 500 ml $ 18.18 CH 2132 15527-013 Glycine 500 g $ 23.53 TC 1182 11605-094 Grace's/Hink's supp - yeastolate+lactalbumin 500 ml $ 16.22 CH 2133 15502-016 Guanidine HCL 500 g $ 65.42 TC 1131 11550-043 Ham's F-10 500 ml $ 6.54 TC 1132 11765-047 Ham's F-12 1L $ 8.56 TC 1231 14175-079 Hank's BSS (1X) w/o Ca++ or Mg++ or phenol 1L $ 13.73 TC 1296 14170-112 HBSS 500 ML $ 8.09 CH 4445 15630-080 HEPES Buffer Solution (1M) 100 ml $ 26.77 RE 6162 25212-010 Hha I 1500u $ 49.45 RE 6169 15207-012 Hind III 5000u $ 14.06 SM 7123 16050-122 Horse Serum 500 ml $ 24.42 MB 4135 15279-011 Human Cot-1® DNA 500 µg $ 108.53 TC 1297 31980-030 IMDM w/GlutaMAX 500 ML $ 6.98 CH 6092 Q602-20 imMedia™ Amp Blue ea $ 225.64 CH 6094 Q612-20 imMedia™ Kan Blue ea $ 177.96 CH 6095 Q621-20 imMedia™ Zeo Agar ea $ 276.96 TC 1147 12440-053 Iscove's Medium 500 ml $ 6.44 OE 5120 18012-021 Klenow - Sequencing Grade 100u $ 39.22 RE 6339 15232-036 Kpn I 5000u $ 37.25 MR 4136 15612-013 Lambda DNA/HindIII Fragment 500 µg $ 90.41 MR 4307 10382-018 Lambda DNA/HindIII Fragment - RTS - ready load 100 app $ 44.63 CH 4186 22700-025 LB Agar 500 g $ 47.15 TC 1187 25030-081 L-Glutamine - 100X - 200mM - 29.23mg/ml 100 ml $ 4.73 MB 6031 11668-019 LipofectAMINE 2000 1.5ml $ 273.82 MB 6032 11668-027 LipofectAMINE 2000 0.75ml $ 166.04 MB 4181 18324-012 Lipofectamine™ 1 ml $ 165.28 MB 4582 18324-020 Lipofectamine™ 4 x 1 ml $ 543.86 MB 4624 18324-111 Lipofectamine™ .5 ml $ 98.67 MB 4138 18292-011 Lipofectin® Reagent 1 ml $ 125.82 MR 5182 LC5600 MagicMark Western Protein Standard 250ul $ 177.71 MR 4014 LC5602 MagicMark XP 250ul $ 178.10 OE 5127 28025-013 Maloney-MLV Reverse Transcriptase 40000u $ 141.01 MR 4596 LC5677 Mark12™ Wide Range Molecular Weight Standard 1.0 ml $ 92.63 CE 5458 12034-013 Max Efficiency DH5@T1 Competent Cells 1 ml (5 x 0.2ml) $ 123.50 CE 5425 18258-012 MAX Efficiency DH5alpha Efficiency Comp Cells 5x.1ml $ 157.71 CE 5474 10268-019 Max Efficiency STBL2 1 ML $ 148.43 TC 1183 16600-082 McCoy's 5A 500 ml $ 4.55 TC 1184 11150-059 Medium 199 500 ml $ 6.86 TC 1154 11095-072 MEM - w/ Earles' salts & L-glut 1L $ 6.59 TC 1230 12571-048 MEM alpha - w/ nucleosides 1L $ 27.27 TC 1155 12561-049 MEM alpha - w/o nucleosides 1L $ 12.63 TC 1156 12561-056 MEM alpha - w/o nucleosides 500 ml $ 7.48 CO 5220 K1520-02 Micro-FastTrack™ Kit 20 rxn $ 369.52 MR 4597 LC5725 MultiMark® Multi-Colored Standard 500 µl $ 106.52 MB 4620 17502-048 N2 Supplement 5 ml $ 38.68 TC 1209 17504-044 Neural Supplement - B27 (50X) 10 ml $ 42.63 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 94 TC 1208 21103-049 Neuralbasal Media 500 ml $ 25.64 KT 5133 FM 2441 18160-010 Nick translation Kit 50 rxn $ 128.09 LC2000 Nitrocellulose Membrane (0.2 µm) Filter Paper Sandwich 20/pk $ 104.21 FM TC 2442 LC2001 Nitrocellulose Membrane (0.45 µm) Filter Paper Sandwich 20/pk $ 104.21 1158 11140-050 Non-essential amino acids - 100X 100 ml $ RE 5.40 6208 15441-025 Not I 200u $ 30.36 CH 4011 NP00061 NP Transfer Buffer (20X) 1L $ 100.04 PG 4013 NP0342 NuPAGE 12% Bis-Tris Gels 1.0mm 12 well ea $ 9.73 PG 4012 NP0335 NuPAGE 4-12% Bis-Tris Gels 1.5mm 10 well EA $ 9.73 PG 6073 NP0301 NuPAGE® 10% Bis-Tris Gels 1.0 mm 10 well ea $ 9.73 PG 6074 NP0302 NuPAGE® 10% Bis-Tris Gels 1.0 mm 12 well ea $ 9.73 PG 6075 NP0303 NuPAGE® 10% Bis-Tris Gels 1.0 mm 15 well ea $ 9.73 PG 6076 NP0341 NuPAGE® 12% Bis-Tris Gels 1.0 mm 10 well ea $ 9.73 PG 6077 NP0343 NuPAGE® 12% Bis-Tris Gels 1.0 mm 15 well ea $ 9.73 PG 6078 NP0321 NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 10 well ea $ 9.73 PG 6079 NP0322 NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 12 well ea $ 9.73 PG 6080 NP0323 NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 15 well ea $ 9.73 CH 6081 NP0005 NuPAGE® Antioxidant 15ml $ 15.28 CH 6082 NP0007 NuPAGE® LDS Sample Buffer (4X) 10ml $ 7.64 CH 6083 NP0002 NuPAGE® MES SDS Running Buffer (20X) 500ml $ 43.07 CH 6084 NP0001 NuPAGE® MOPS SDS Running Buffer (20X) 500ml $ 43.07 CH 6085 NP0004 NuPAGE® Sample Reducing Agent (10X) 250ul $ 7.64 CH 6086 NP0006 NuPAGE® Transfer Buffer (20X) 125ml $ 13.43 CH 6087 LA0041 NuPAGE® Tris-Acetate SDS Running Buffer (20X) 500ml $ 41.22 MB 6125 18418-012 Oligo (dT) 12-18 Primer 25 UG $ 45.40 MB 4009 12252-011 Oligofectamine Reagent 1 ML $ 188.12 CE 4637 C862003 One Shot MACH 1 Comp eColi (LTI Shelf) 20 rxn $ 235.31 CE 4622 12297-016 One Shot® MAX Efficiency® DH5a®-T1® 20 x 50ul $ 243.98 CE 4334 C2020-03 OneShot™ INVaF' Competent Cells 21x50µl $ 243.61 CE 5445 C4040-03 OneShot™ TOP10 Competent Cells 21x50µl $ 238.08 CE 4335 C3030-03 OneShot™ TOP10F' Competent Cells 21x50µl $ 235.31 TC 1190 09400-72PF OPI 100X - 15mg O - 5 mg P - 20u I/ml 50 ml $ 24.13 TC 1163 31985-070 Opti-MEM® - reduced serum hybrid. medium 500 ml $ 14.29 KT 6098 K4900-01 pcDNA3.1© Directional TOPO® Expression Kit (Top 10 Cells) 20 rxn $ 360.57 TC 1188 15140-122 Pen/Strep - 100X -10000u/ml P - 10mg/ml S 100 ml $ 4.47 TC 1169 12040-077 PFHM II - Protein Free Hybridoma Medium 1L $ 34.96 TC 1170 12040-093 PFHM II - Protein Free Hybridoma Medium 500 ml $ 20.39 CH 2176 15513-039 Phenol Saturated (Buffer) 100 ml $ 58.24 MR 4151 15611-015 Phi-X174 DNA/Hae III Fragment 40 µg $ 54.30 MR 4305 10383-016 Phi-X174 DNA/Hae III Fragment - RTS - ready load 100 app $ 83.60 PC 6052 11708-013 Platinum Pfx DNA Polymerase 100 u $ 81.62 PC 6053 11708-021 Platinum Pfx DNA Polymerase 250 u $ 185.54 PC 6054 11708-039 Platinum Pfx DNA Polymerase 500 u $ 350.35 PC 5459 10790020 Platinum Supermix Hi Fi 100 rxns $ 121.70 PC 6013 10966-018 Platinum Taq DNA Polymerase - 100u 100u $ 46.05 PC 6014 10966-026 Platinum Taq DNA Polymerase - 250u 250u $ 110.99 PC 6015 10966-034 Platinum Taq DNA Polymerase - 500u 500u $ 200.72 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 95 PC 5469 11304-011 Platinum Taq High Fidelity 100u $ MB 6034 11514-015 Plus Reagent .85 ml $ 87.73 IM 3111 15918-014 Protein A Agarose 5 ml $ 166.69 IM 3112 15920-010 Protein G Agarose 5 ml $ 253.01 FM 2443 LC2002 PVDF Membrane/Filter Paper Sandwich 20/pk $ 101.89 KT 5227 18428-011 Rad Prime Labeling Kit 30 rxn $ 128.88 MR 4632 12308-011 Ready Load 1KB Plus DNA Ladder 100 rxn $ 73.79 OE 5141 18021-014 Ribonuclease H 30u $ 73.88 MR 4172 15620-016 RNA Ladder 0.24-9.5 kb 75 µg $ 80.29 TC 1171 11875-085 RPMI 1640 1L $ 6.17 TC 1226 11835-030 RPMI 1640 - no phenol 500 ml $ 4.90 TC 1173 22400-089 RPMI 1640 - w/ 25mM Hepes 500 ml $ 6.14 TC 1299 72400-047 RPMI w/GlutaMAX 500 ML $ 6.45 TC 1210 15544-018 S.O.C. Transformation Media - 10x10 ml 100 ml $ 43.41 CH 2284 15553-027 SDS 10% (4x100ml) 400 ml $ 25.51 MR 4599 LC5925 SeeBlue® Plus2 Pre-Stained Standard 500 µl $ 99.11 MR 4598 LC5625 SeeBlue® Pre-Stained Standard 500 µl $ 99.11 TC 1185 10902-096 SF900 II - Serum-free insect medium 500 ml $ 19.17 TC 1277 10902-088 SF900 II - Serum-free insect medium 1L $ 41.98 MR 4600 LC6060 SimplyBlue™ SafeStain 1L $ 79.66 RE 6286 15228-018 Sma I 1000u $ 44.28 TC 1235 25080-094 Sodium Bicarbonate 100 ml $ 4.04 OE 5156 18018-010 Sp6 RNA Polymerase 500u $ 48.07 RE 6256 15413-016 Sph I 200u $ 64.44 CH 2279 15557-036 SSC 20X 4L $ 49.53 CH 2285 15591-035 SSPE 20X 4L $ 79.75 RE 6261 15222-011 Sst I 2000u $ 71.60 IM 3116 19543-024 Streptavidin Red 670™ 100 µg $ 157.50 OE 5460 11917010 Superscript Custom Choice Kit 10 rxn $ 656.60 KT 5226 11904-018 Superscript First-Strand Synthesis System for RT-PCR 50 rxn $ 235.30 OE 5478 18080044 Superscript III Reverse Transcriptase 10000u $ 158.13 OE 5476 12574-026 SuperScript III, 1 step w/Platinum Taq 100 100 rxn $ 351.55 PC 5467 10928-034 SuperScript One Step RT-PCR with Platinum Taq 25 rxn $ 117.78 OE 5159 18064-014 Superscript™ II Reverse Transcriptase 10000u $ 149.91 PC 5465 18064-071 Superscript™ II Reverse Transcriptase 4 x 10000u $ 599.63 OE 5160 18053-017 Superscript™ Reverse Transcriptase 10000u $ 154.90 OE 5164 15224-017 T4 DNA Ligase 100u $ 26.09 OE 5229 15224-025 T4 DNA Ligase 500u $ 115.12 OE 5168 18005-017 T4 DNA Polymerase 50u $ 31.89 OE 5464 18005-025 T4 DNA Polymerase 250u $ 103.79 OE 5172 18004-010 T4 Polynucleotide Kinase 200u $ 50.03 KT 5389 K2050-01 TA Cloning Kit® with Dual Promoters (INVaF') 20 rxn $ 289.93 CH 2275 15558-026 TAE 10X 4L $ 58.24 PC 5187 18038-018 Taq DNA Polymerase w/ buffer 100u $ 29.01 PC 5188 18038-042 Taq DNA Polymerase w/ buffer 500u $ 115.86 OE 5194 10533-065 Terminal Deoxy Transferase (TdT) 500u $ 108.42 KT 6097 K4500-01 TOPO TA Cloning® Kit (TOP10 Cells) 20 rxn $ 322.35 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 96 102.47 KT 5397 K4550-01 TOPO TA Cloning® Kit (TOP10F' Cells) 20 rxns $ 308.67 KT 5466 K4520-01 TOPO TA Cloning® Kit (with LTI 4622 - DH5a Cells) 20 rxns $ 319.57 KT 5398 K4650-01 TOPO TA Cloning® Kit Dual Promoter (TOP10F' Cells) 1 ea $ 322.99 KT 5446 K4575-01 TOPO TA Cloning® Kit for Sequencing (Top 10 Cells) 20 rxn $ 334.39 CH 2281 15567-027 TRIS pH 7.5 1L $ 17.73 CH 2282 15568-025 TRIS pH 8.0 1L $ 19.28 CH 2202 15506-017 Tris Hydrochloride 500 g $ 50.10 CH 5457 15596018 Trizol 200 ml $ 149.82 MB 4182 15596-026 TRIzol Reagent 100 ml $ 78.80 TC 1232 15090-046 Trypsin - 2.5% (10X) 100 ml $ 14.29 TC 1176 25300-054 Trypsin-EDTA - 0.05% T (1X) 100 ml $ 3.50 TC 1236 25200-056 Trypsin-EDTA - 0.25% (1X) 100 ml $ 5.48 TC 1177 15400-054 Trypsin-EDTA - 0.5% (10X) 100 ml $ 6.27 CH 2204 15505-050 Urea 2 kg $ 67.36 TC 1179 11120-052 Vitamin Mixture - 100X vitamin supplement 100 ml $ 6.84 RE 6269 15226-012 Xba I 2000u $ 26.26 RE 6344 15226-038 Xba I 10000u $ 113.20 MB 4180 15401-011 Yeast tRNA 25 mg $ 44.91 CH 4189 22712-020 YT Broth 2X 500 g $ 33.58 MB 5417 R250-01 Zeocin 1g $ 175.07 KT 6096 K2800-20 Zero Blunt® TOPO® PCR Cloning Kit (TOP 10 Cells) 20 rxn $ 338.20 KODAK .............................................................................................................KOD _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION FM 2306 V8701302 Biomax™ Film MR-1 8x10in FM 2307 V8715187 Biomax™ Film MR-1 35x43cm FM 2364 V8294985 Biomax™ Film MS-1 8x10in FM 2382 EB135-24 Elite II Slide Film 100 - 24 Exposures FM 2383 EB135-36 Elite II Slide Film 100 - 36 Exposures FM 2331 V1651678 X-Omat™ AR-2 interleaved film 35x43cm FM 2332 V1651579 X-Omat™ AR-2 interleaved film 8x10in FM 2211 V1651454 X-Omat™ AR-5 non-interleaved film 8x10in SIZE 50 sht 50 sht 50 sht 1 roll 1 roll 50 sht 50 sht 50 sht CC PRICE $ 138.93 $ 300.20 $ 118.70 $ 6.76 $ 9.13 $ 353.46 $ 182.70 $ 144.05 MARLIGEN BIOSCIENCES, INC. ………………………………………………………………………MLG _ Technical Services: (866) 464-4990 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: No _ Delivery: 2nd Business Day PC CODE MFGR. NO. DESCRIPTION KT 5622 11452-018 High Pure Maxi Kit KT 5625 11456-019 Rapid Gel Extraction Kit KT 5624 11458-015 Rapid PCR Purification Kit KT 5623 11453-016 Rapid Plasmid Mini Prep SIZE 10rxn 50rxn 50rxn 50rxn CC PRICE. $ 122.65 $ 54.28 $ 54.28 $ 48.83 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 97 MEDIATECH (FISHER) ........................................................................................................ MED _ Technical Services: (800) CELLGRO _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: No _ Delivery: 3rd Business Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE CH 1290 MT46-010-CM 10X TAE 1L $ CH 1291 MT46011CM 10X TBE 1L $ CH 1292 MT46020CM 20X SSC Buffer 1L $ TC 1273 MT10013CV DMEM - high glucose 4.5 gm/L 500 ml $ TC 1274 MT10017CV DMEM - high glucose 4.5 gm/L - no sodium pyruvate 500 ml $ TC 1239 MT10013CM DMEM WITH L-GLUT; HI GLUCOSE; W/ NA PYRUVATE 1 L $ TC 1260 MT10017CM DMEM WITH L-GLUT; HI GLUCOSE; W/O NA PYRUVATE 1 L $ TC 1241 MT21030CM DPBS (1X) with Ca++ & Mg++ 1L $ TC 1243 MT21031CM DPBS (1X) without Ca++ & Mg++ 1L $ TC 1264 MT61234RF G418 1g $ TC 1265 MT61234RG G418 5g $ TC 1282 MT30-234-CR G418 Sulfate (50mg/ml) 20 ml $ TC 1245 MT10080CV Ham's F-12 500 ml $ TC 1275 MT21020CM HBSS w/ ca and mg 1L $ TC 1276 MT21021CM HBSS w/o ca and mg 1L $ TC 1283 MT30-240-CR Hygromycin B (50mg/ml) 20 ml $ TC 1247 MT10016CV Iscove's Medium 500 ml $ CH 1294 MT46-050-CM LB BROTH 1L $ TC 1249 MT25005CI L-Glutamine - 200mM 100 ml $ TC 1251 MT10010CM MEM - w/ Earles' salts & L-Glut 1L $ TC 1253 MT30002CI Pen/Strep - 100X -10mcg/ml P - 10mg/ml S 100 ml $ TC 1255 MT10040CM RPMI 1640 1L $ TC 1256 MT10041CV RPMI 1640 - w/ 25mM Hepes 500 ml $ TC 1295 MT25-000-CI Sodium Pyruvate, 100mM solution 100 ml $ CH 1293 MT25-900-CI Trypan Blue Solution 100ML $ TC 1259 MT25052CI Trypsin-EDTA - 0.05% T - .53mM EDTA 100 ml $ MILLIPORE.......................................................................................MIL _ Technical Services: (800) 225-1380 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 2nd Business Day PC CODE MFGR. NO. DESCRIPTION FM 2439 INYC13750 Immobilon-NY+ 137mm DISC(Colony Plaque) FM 2214 IPVH20200 Immobilon™ P 20x20cm (Western Blot) FM 2215 IPVH00010 Immobilon™ P 26.5cmx3.75m (Western Blot) FM 2217 HATF08250 Nitrocellulose 82mm Colony & Plaque FM 2216 HATF13750 Nitrocellulose 137mm Colony & Plaque FM 2271 SCGPU02RE Stericup Filtr. & Stor.-PES 250ml .22µ FM 2272 SCGPU05RE Stericup Filtr. & Stor.-PES 500ml .22µ FM 2273 SCGPU11RE Stericup Filtr. & Stor.-PES 1000ml .22µ FM 2433 SCGPU01RE Stericup Filtr. & Stor.-PES 150ml .22µ FM 2339 SCGP00525 Steriflip-GP Filter Unit 50µl FM 2434 SCGPS10RE Steritop-GP Filtr 1000 ML 33 mm neck 0.22µm FM 2435 SCGPT10RE Steritop-GP Filtr 1000 ML 45 mm neck 0.22µm FM 2336 SCGPS05RE Steritop-GP Filtr 500 ML 33 mm neck 0.22µm FM 2335 SCGPT05RE Steritop-GP Filtr 500 ML 45 mm neck 0.22µm SIZE 50/pk 10/pk 1 roll 50/pk 50/pk 12/pk 12/pk 12/pk 12/pk 25/pk 12/pk 12/pk 12/pk 12/pk CC PRICE $ 156.48 $ 101.81 $ 125.90 $ 49.63 $ 101.81 $ 42.64 $ 66.82 $ 95.45 $ 33.09 $ 80.25 $ 65.54 $ 65.54 $ 41.36 $ 41.36 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 98 12.70 11.65 9.51 3.98 3.98 7.96 7.96 7.96 7.96 58.11 201.69 63.10 4.24 7.96 7.96 97.69 6.37 13.78 4.77 7.96 4.77 7.96 5.84 4.30 6.63 4.78 FM FM FM FM 2266 2264 2267 2265 SLGV025LS SLHVO25LS SLGS0250S SLHA0250S Syringe fltr-Durapore PVDF/PVC .22µ Syringe fltr-Durapore PVDF/PVC .45µ Syringe fltr-mixed esters of cellulose .22µ Syringe fltr-mixed esters of cellulose .45µ 50/pk 50/pk 50/pk 50/pk $ $ $ $ 70.63 66.49 64.27 62.36 SIZE 50/pk 50/pk 10/pk 50/pk 1 roll 1 roll 50/pk 1 roll CC PRICE $ 107.32 $ 125.41 $ 23.91 $ 104.99 $ 99.17 $ 146.88 $ 50.75 $ 140.31 MICRON SEPARATIONS INC. (FISHER) .............................................................................. MSI _ Technical Services: (800) 444-8212 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 3rd Business Day PC CODE MFGR. NO. DESCRIPTION FM 2432 N00HN13750 Magna 137mm Notched FM 2150 NJ0HY13750 Magnagraph 137mm FM 2428 EP4HY408F0 Nitrobind 7x8.4 FM 2157 EP4HY13750 Nitrobind Pure 137mm FM 2158 EP4HYA0010 Nitrobind Pure 20cmx3m FM 2159 EP4HY00010 Nitrobind Pure 30cmx3m FM 2164 WP4HY08250 Nitropure Supp 82mm FM 2163 WP4HY00010 Nitropure Supp 30cmx3m NATIONAL DIAGNOSTICS.................................................................ND _ Technical Services: (800) 526-3867 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 1 - 2 Weeks PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE CH 2101 EC850 AccuGel™ 19:1 450 ml $ CH 2177 EC890 ProtoGel® (30% Acrylamide/0.8% Bis) 1L $ CH 2341 EC-892 ProtoGel® Buffer 450 ml $ CH 2342 EC-893 ProtoGel® Stacking Buffer 200 ml $ CH 2179 EC835 SequaGel® Buffer 200 ml $ CH 2180 EC830 SequaGel® Concentrate (20% Acryl.) 19:1 1L $ CH 2181 EC840 SequaGel® Diluent 1L $ CH 2182 EC833 SequaGel® Sequencing System 1 kit $ CH 2184 EC836 SequaGel®-6 6% Monomer Sequencing Soln (includes EC-841) 1 L $ CH 2183 EC838 SequaGel®-8 8% Monomer Sequencing Soln (includes EC-841) 1 L $ NEW ENGLAND BIOLABS ...................................................................................................NEB _ Technical Services: (800) 632-5227 _ Website: http://www.neb.com/ _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: All items in the New England Biolabs catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders for NEB products must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Monday. _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE MR 4584 N3232S 1 kb DNA Ladder 100ug $ 40.44 MR 4583 N3231S 100 bp DNA Ladder 50ug $ 40.44 RE 6101 R0117S Aat II 500u $ 36.77 RE 6104 R0161S Acc I 500u $ 44.12 RE 6107 R0520S Afl II 2000u $ 36.77 RE 6312 R0541S Afl III 250u $ 40.44 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 99 43.76 72.98 19.36 16.00 10.00 54.20 20.90 85.58 37.44 39.98 OE RE RE MB RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE OE RE RE OE RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE OE RE MR MR MB RE RE RE RE RE RE RE RE RE 5307 6410 6414 4330 6111 6288 6323 6116 6385 6416 6119 6297 6124 6128 6298 6408 6299 6387 6381 6137 6384 6388 5110 6139 6142 5112 6140 6389 6390 6143 6300 6147 6301 6152 6159 6325 6166 6303 6176 6180 6182 5122 6185 4252 4137 4331 6190 6324 6123 6413 6193 6305 6411 6201 6204 M0392S R0552S R0514S E8021L R0114S R0558S R0526S R0153S R0174S R0174L R0136S R0136L R0136T R0160S R0144S R0144M R0540S R0517S R0199S R0162S R0113S R0524S M0290S R0197S R0175S M0209S R0176S R0129S R0510S R0505S R0521S R0101S R0101L R0101T R0107S R0139S R0103S R0104S R0155S R0105S R0171S M0210S R0142S N3011S N3012S S1404S R0147S R0589S R0198S R0163S R0534S R0190S R0191S R0193S R0111S Agarase Age I AlwN I Amylose Resin Apa I Asc I Ase I Ava II Avr II Avr II BamH I BamH I BamH I HC Bcl I Bgl II Bgl II BspE I BspH I BssH II BstE II BstX I Bsu36 I Calf Intestinal Alkaline Phosphatase Cla I Dde I DNA Polymerase I from E.coli Dpn I Dra I Dra III Eag I (Xma III) EcoN I EcoR I EcoR I EcoR I HC Hae II Hha I Hinc II Hind III Hinf I Hpa I Hpa II Klenow - Sequencing Grade Kpn I Lambda DNA Lambda DNA/HindIII Fragment m7GpppG 5'G RNA Cap Structure Mbo I Mfe I Mlu I Mnl I Msc I Nae I Nar I Nco I Nde I 100u 250u 500u 100 ml 2000u 500u 2000u 2000u 100u 500u 10000u 50000u 10000u 3000u 2000u 10000u 1000u 500u 200u 2000u 1000u 500u 1000u 1000u 500u 500u 1,000u 2000u 1000u 500u 1000u 10000u 50000u 10000u 2000u 2000u 1000u 10000u 5000u 500u 2000u 200u 3000u 250 µg 150 µg 25u 500u 500u 1000u 250u 100u 500u 200u 1000u 4000u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 44.12 40.44 40.44 220.61 44.12 40.45 40.45 36.77 44.12 176.48 36.77 147.07 36.77 36.77 36.77 147.07 40.44 40.44 40.44 36.77 40.44 40.44 40.44 40.44 36.77 40.44 40.44 40.44 40.44 40.44 40.44 36.77 147.07 36.77 36.77 36.77 36.77 36.77 36.77 36.77 36.77 36.77 36.77 40.44 40.44 110.31 44.12 44.13 40.36 40.44 40.44 36.77 36.77 36.77 40.44 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 100 RE RE RE RE RE RE RE MB MR MR RE MR RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE RE OE OE OE OE OE OE PC PC RE RE RE RE RE RE 6205 6308 6210 6309 6213 6214 6409 4254 4250 4251 6412 4546 6220 6227 6230 6234 6314 6236 6315 6322 6241 6246 6249 6252 6254 6258 6392 6263 6383 5165 5217 5166 5169 5173 5218 5257 5195 6271 6320 6274 6287 6393 6280 R0131S R0131L R0189S R0189L R0192S R0127S R0547S N3032S N3026L N3026S R0560S P7708S R0140S R0151S R0167S R0156S R0157S R0138S R0138L R0569S R0169S R0123S R0141S R0130S R0133L R0182S R0132S R0187S R0500S M0202S M0202L M0202T M0203S M0201S M0201L M0257S M0254S R0145S R0145L R0146S R0146L R0180S R0194S Nhe I Nhe I Not I Not I Nru I Nsi I Pac I pBR322/Msp I Digest Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Pme I Prestained Protein Molecular Weight Marker Pst I Pvu II Rsa I Sac I (Sst I) Sac II Sal I Sal I Sap I Sau 3A I Sfi I Sma I SnaB I Spe I Sph I Ssp I Stu I Sty I T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase HC T4 DNA Polymerase T4 Polynucleotide Kinase T4 Polynucleotide Kinase Vent® (exo-) Polymerase Vent® DNA Polymerase Xba I Xba I Xho I Xho I Xma I Xmn I 1000u 5000u 500u 2500u 1000u 1000u 250u 50 µg 250u 50u 500u 175 lns 10000u 5000u 1000u 2000u 2000u 2000u 10000u 50u 200u 2000u 2000u 500u 2500u 250u 1000u 1000u 3000u 20000u 100000u 20000u 150u 500u 2500u 200u 200u 3000u 15000u 5000u 25000u 250u 1000u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 40.44 161.78 44.12 176.48 36.77 40.44 40.44 40.44 161.78 40.44 44.12 62.51 40.44 36.77 36.77 36.77 36.77 36.77 147.07 40.45 36.77 40.44 36.77 36.77 161.78 40.44 40.44 36.77 44.12 44.12 176.48 44.12 40.44 36.77 147.07 58.83 58.83 44.12 176.48 44.12 176.00 40.44 36.77 PERKIN ELMER (NEW ENGLAND NUCLEAR) ................................................................... NEN _ Technical Services: (800) 632-5227 _ Web Site: www.perkinelmer.com _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: Many non-radioactive items in the NEN catalog may be special ordered, no shipping or dry ice fees. _ Items ordered on Friday will be available on Tuesday. _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 101 PC PC NR FM FM FM FM FM NR NR NR NR FM FM 5455 5456 5979 2260 2250 2246 2249 2253 5201 3120 3121 3122 2388 2385 NEL542 NEL999 NEL-601 NEF-978Y NEF-986 NEF-976 NEF-984 NEF-1002 NEL-201 NEL-103 NEL-104 NEL-105 NEF-586 NEF-596 Biotin 11 CTP 250nm Biotin 16 UTP 250nm CDP-Star Nucleic Acid Chemiluminescence Reagent (Ready to Use) 1 kit Colony/Plaque Screen™ w/orient. holes 137mm 50/bx Genescreen Plus® Hybridization Mem. 20x20cm 10/bx Genescreen Plus® Hybridization Mem. 30x45cm 5 sht Genescreen™ Hybridization Mem. 20x20cm 10/bx Polyscreen® PVDF 30cmx3m 1 roll Renaissance™ Nucleic Acid Chemilum Rgt Kit 1 kit Renaissance™ Western Blot Chemilum Rgt 1000cm2 Renaissance™ Western Blot Chemilum Rgt 2500cm2 Renaissance™ Western Blot Chemilum Rgt 5000cm2 X-Omat™ Blue 5x7 100/bx X-Omat™ Blue 8x10in 100/bx $ $ $ $ $ $ $ $ $ $ $ $ $ $ APPLIED BIOSYSTEMS ....................................................................................................... PEC _ Technical Services: (800) 762-4001 _ Web Site: http://www.appliedbiosystems.com/ _ All items listed below are available on site at the Cell Center. _ Special Ordering: All items in the PEC catalog may be special ordered at a discount, no shipping or dry ice fees. _ Conditions: All special orders for PEC products must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Monday. _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE PC 5993 N808-0246 AmpliTaq Gold™ w/ buffer I - 1000u 1000u $ 284.65 PC 5382 N808-0240 AmpliTaq Gold™ w/ buffer I - 250u 250u $ 86.08 PC 5997 N808-0244 AmpliTaq Gold™ w/ buffer I-12 pac (250u) 3000u $ 1,032.97 PC 5999 N808-0248 AmpliTaq Gold™ w/ buffer I-5 pac (1000u) 5000u $ 1,413.18 PC 5995 N808-0242 AmpliTaq Gold™ w/ buffer I-6 pac (250u) 1500u $ 516.49 PC 5994 N808-0247 AmpliTaq Gold™ w/ buffer II - 1000u 1000u $ 284.65 PC 5383 N808-0241 AmpliTaq Gold™ w/ buffer II - 250u 250u $ 86.08 PC 5998 N808-0245 AmpliTaq Gold™ w/ buffer II-12 pac (250u) 3000u $ 1,032.97 PC 6000 N808-0249 AmpliTaq Gold™ w/ buffer II-5 pac (1000u) 5000u $ 1,413.18 PC 5996 N808-0243 AmpliTaq Gold™ w/ buffer II-6 pac (250u) 1500u $ 516.49 PC 5263 N808-0171 AmpliTaq® w/ buffer I - 1000u 1000u $ 288.37 PC 5264 N808-0152 AmpliTaq® w/ buffer I - 12 paq (250u) 3000u $ 900.62 PC 5212 N808-0160 AmpliTaq® w/ buffer I - 250u 250u $ 75.05 PC 5265 N808-0155 AmpliTaq® w/ buffer I - 5 paq (1000u) 5000u $ 1,431.72 PC 5266 N808-0166 AmpliTaq® w/ buffer I - 6 paq (250u) 1500u $ 450.32 PC 5267 N808-0172 AmpliTaq® w/ buffer II - 1000u 1000u $ 288.37 PC 5213 N808-0161 AmpliTaq® w/ buffer II - 250u 250u $ 75.05 PC 5268 N808-0156 AmpliTaq® w/ buffer II - 5 paq (1000u) 5000u $ 1,431.72 PC 5269 N808-0167 AmpliTaq® w/ buffer II - 6 paq (250u) 1500u $ 450.32 PC 5270 N808-0153 AmpliTaq® w/ buffer II- 12 paq (250u) 3000u $ 900.62 PC 5251 N808-0150 AmpliWax® PCR Gem 50 200 gem $ 73.50 PC 5250 N808-0100 AmpliWax® PCR Gem 100 200 gem $ 73.50 PC 5407 4303573 BigDye™ Terminator Kit 24 rxn $ 192.37 PC 5404 N808-0260 GeneAmp® dNTP blend 10mM 1 ml $ 29.40 PC 5406 4303442 GeneAmp® dNTP blend 10mM-12 paq 12 ml $ 352.80 PC 5405 4303441 GeneAmp® dNTP blend 10mM-6 paq 6 ml $ 176.41 PC 5191 N808-0009 GeneAmp® Core Kit 100 rxn $ 164.21 PC 5247 N808-0007 GeneAmp® dNTP Set -10mM (4x3.2umol) 1 set $ 36.50 PC 5253 N808-0179 GeneAmp® EZ rTth RNA PCR Kit 100 rxn $ 355.64 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 102 187.07 186.03 134.25 231.21 150.24 218.43 149.17 181.14 131.06 77.79 140.65 198.18 57.53 135.27 PC PC PC PC PC PC PC PC PC 5202 5248 5255 5254 5249 4628 4629 4626 4627 N808-0143 N808-0193 N808-0178 N808-0098 N808-0180 4309155 4312704 4304437 4305719 GeneAmp® RNA PCR Core Kit GeneAmp® XL PCR Kit rTth DNA Polymerase & EZ Buffer Pack rTth DNA Polymerase with Buffer rTth DNA Polymerase-XL w/XL Buffer Pack Sybr Green PCR Master Mix Sybr Green PCR Master Mix TaqMan Universal Master Mix TaqMan Universal Master Mix 100 rxn 100 rxn 100 rxn 100 rxn 100 rxn pk 10 pk 200 rxn 2000 rxn $ $ $ $ $ $ $ $ $ 237.21 375.40 229.19 229.19 190.34 273.52 2,679.00 310.00 3,035.00 POLAROID ...............................................................................................................POL _All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: NO _ Delivery: 3rd Business Day PC CODE MFGR. NO. DESCRIPTION FM 2322 444159 Polaroid Film Type 57 FM 2323 444191 Polaroid Film Type 667 SIZE pk pk CC PRICE $ 47.95 $ 24.07 PROMEGA...........................................................................................PRO _ Technical Services: (800) 356-9526 _ Web Site: http://www.promega.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: No _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION MR 4588 PRG6941 1 kb DNA Step Ladder MR 4585 G4471 10 bp DNA Step Ladder MR 4455 G2101 100 bp DNA Ladder MR 4586 G6951 100 bp DNA Step Ladder MR 4587 G6961 200 bp DNA Step Ladder MR 4590 G4521 50 bp DNA Ladder OE 5105 M5108 AMV Reverse Transcriptase OE 5106 M5101 AMV Reverse Transcriptase RE 6396 R6361 Apa I RE 6121 R6021 BamH I RE 6398 R6071 Bgl I RE 6133 R6081 Bgl II RE 6400 R6641 BstE II OE 5256 M1821 Calf Intestinal Alkaline Phosphatase RE 6289 R6551 Cla I PC 4241 U1201 dATP (100mM) PC 4242 U1221 dCTP (100mM) RE 6401 R6291 Dde I PC 4243 U1211 dGTP (100mM) OE 5204 M2051 DNA Polymerase I OE 5144 M6101 DNase RQ1 RNase-free PC 4240 U1240 dNTP's (bulk) (100mM) 4x40umoles PC 4244 U1231 dTTP (100mM) RE 6149 R6011 EcoR I RE 6290 R6351 EcoR V OE 5262 M1811 Exonuclease III MB 4248 Q4100 fmol® DNA Sequencing System CE 4246 L2011 HB101 High Efficiency Competent Cells SIZE 300 lns 50 µl 250 µl 100 µl 100 µl 100 lns 1000u 300u 5000u 2500u 1000u 500u 2000u 1000u 500u 40 µmol 40 µmol 200u 40 µmol 500u 1000u 1 set 40 µmol 5000u 2000u 5000u 1 kit 1 ml CC PRICE $ 58.57 $ 57.08 $ 56.38 $ 75.92 $ 78.51 $ 65.48 $ 131.21 $ 50.41 $ 38.32 $ 24.47 $ 30.95 $ 22.24 $ 38.32 $ 35.58 $ 35.58 $ 61.74 $ 61.74 $ 17.81 $ 61.74 $ 66.72 $ 22.19 $ 193.18 $ 61.74 $ 17.80 $ 31.14 $ 45.70 $ 143.73 $ 58.83 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 103 RE RE RE CE OE OE RE MR NR NR NR OE RE RE RE RE RE RE RE MB PC MB MB MB MR CO RE RE RE MB OE MB MB MB MB OE RE RE RE RE RE RE RE OE RE RE RE RE RE OE OE OE OE OE PC 6358 6172 6361 4247 5366 5123 6363 4486 3185 3212 3211 5367 6364 6365 6203 6367 6206 6292 6368 4268 4199 5942 5943 4294 4198 4528 6369 6370 6371 4169 5205 4176 4177 4174 4175 5207 6373 6374 6238 6375 6243 6251 6376 5158 6293 6260 6403 6377 6404 5161 5208 5209 5175 5179 5982 R6441 R6041 R6301 L2001 M2181 M2201 R6341 G1711 E4030 E1501 E4530 M1701 R6711 R6381 R6513 R6801 R6501 R6431 R7091 C1101 PRG3161 A1360 A1380 A3600 G1761 Z5400 R6111 R6321 R6331 L4960 M4281 N2515 N2511 N2115 N2111 M5761 R6061 R6221 R6051 R6191 R6211 R6121 R6791 P1085 R6591 R6261 R6601 R6421 R6481 P2083 M1801 M4211 M4101 P2075 M1868 Hha I Hind III Hpa I JM109 High Efficiency Competent Cells Klenow - Exo Minus Klenow Fragment Kpn I Lambda DNA/HindIII Fragment Luciferase Assay System Luciferase Assay System (10 pk) quote#5075 Luciferase Assay System–10 pack Maloney-MLV Reverse Transcripts Mbo I Mlu I Nco I Nde I Nhe I Not I Nru I Oligo(dT) 15 primer PCR Markers pGEM®-T Easy I pGEM®-T Easy II pGEM®-T Vector I Phi-X174 DNA/Hae III Fragment PolyATtract® System 1000 RNA Isolation System Pst I Pvu I Pvu II Rabbit Reticulocyte Lysate Ribonuclease H RNasin® Recombinant Ribonuclease Inhibitor RNasin® Recombinant Ribonuclease Inhibitor RNasin® Ribonuclease Inhibitor RNasin® Ribonuclease Inhibitor S1 Nuclease Sac I (Sst I) Sac II Sal I Sau 3A I Sca I Sma I SnaB I Sp6 RNA Polymerase Spe I Sph I Ssp I Stu I Sty I T3 RNA Polymerase T4 DNA Ligase T4 DNA Polymerase T4 Polynucleotide Kinase T7 RNA Polymerase Taq DNA Polymerase 1000u 5000u 100u 1 ml 100u 150u 2500u 100µg 100 assays 1000 assays 1000 assays 10000u 200u 1000u 200u 500u 250u 200u 200u 20 µg 250 µl 1 kit 1 kit 20u 50 µg 1 kit 3000u 100u 1000u 1 ml 50u 10000u 2500u 10000u 2500u 10000u 1000u 500u 2000u 100u 1000u 1000u 100u 1000u 200u 200u 500u 400u 2000u 1000u 100u 100u 100u 1000u 2500u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 26.53 14.09 29.48 67.61 27.43 54.87 33.17 24.45 46.84 308.22 412.41 31.88 36.12 35.58 48.92 66.34 40.77 35.58 34.65 33.28 54.12 89.93 151.91 74.41 65.24 279.25 18.53 49.70 22.85 94.41 97.86 173.32 61.49 163.43 64.86 19.27 29.48 37.07 38.55 24.33 35.58 39.07 29.48 94.73 45.21 44.94 49.39 29.48 35.38 22.97 22.98 48.19 27.43 30.39 433.71 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 104 PC PC OE MB KT KT KT KT KT KT IM CO CO CO CO CO CO CO CO CO CO CO CO RE RE 5189 5258 5210 5944 5930 5931 5929 5928 5933 5932 3203 4142 4143 4144 4145 4195 4292 4293 4147 4146 4291 4529 4531 6379 6380 M1861 M1865 M1871 E2381 L4600 L5020 L4950 L4610 L1170 L5010 S3841 PRA7280 PRA7290 PRA7270 A7300 PRA7640 A7211 PRA7141 A7170 A7100 A7510 A1330 A1460 R6181 R6161 Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Terminal Deoxy Transferase (TdT) Tfx-10 Transfection Reagent TNT®-SP6 Coupled Transcription/Translation TNT®-SP6/T7 Coupled Transcription/Translation TNT®-T3 Coupled Transcription/Translation TNT®-T7 Coupled Transcription/Translation TNT®-T7 QUICK Coupled Transcription/Translation TNT®-T7/T3 Coupled Transcription/Translation Western Blue Stabilized Substrate for AP Wizard™ DNA Clean-up System Wizard™ Lambda System Wizard™ Maxipreps System Wizard™ Megapreps System Wizard™ Midipreps System Wizard™ Minicolumns Wizard™ Miniprep DNA Purification Resin Wizard™ PCR Preps System Wizard™ Plus Miniprep System Wizard™ Plus Miniprep System Wizard™ Plus SV Wizard™ Plus SV Xba I Xho I 100u 500u 300u 1 ea 1 kit 1 kit 1 kit 1 kit 1 kit 1 kit 1 ea 100 col 20 col 10 col 5 col 25 col 250 col 250 ml 50 col 50 col 250 col 50 rxn 250 rxn 2000u 3000u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 24.33 82.75 39.29 143.21 291.15 300.09 312.11 293.37 297.05 314.95 30.78 83.87 56.77 90.35 86.36 111.78 99.95 94.35 59.20 65.09 222.20 42.56 189.73 38.55 24.46 QIAGEN ...........................................................................................................QIA _ Technical Services: (800) 426-8157 _ Web Site: http://www.qiagen.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: All items in the QIA catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders for QIA products must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Monday. _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE CO 4603 34698 Anti-His Antibody Selector Kit (3 x 3 µg) 3x3ug $ 70.88 KT 5473 69504 DNeasy Tissue Kit 50 $ 98.69 CO 4601 301425 Effectene Transfection Reagent (1 ml) 1 ml $ 197.38 CO 4502 12362 EndoFree Plasmid Maxi Kit (10) kit $ 211.73 CO 4602 12643 Hispeed Midi Kit (25) 45 minutes/200ug 45min/200ug $ 220.71 KT 5471 12662 Hispeed Plasmid Maxi Kit 10 prep $ 183.02 KT 5472 12663 Hispeed Plasmid Maxi Kit 25 prep $ 442.31 PC 6100 203205 Hot Star Taq (1000) with Q solution (4x250u) $ 261.24 PC 6099 203203 Hot Star Taq (250) with Q solution 250u $ 92.42 CO 4503 30410 Ni-NTA Superflow 25 ml $ 221.60 MS 3345 2003482 Oligo CD - $1,000 EA $ 1,000.00 MS 3344 2003481 Oligo CD - $500 EA $ 500.00 CO 4318 51104 QIAamp™ Blood Kit 50 prp $ 90.61 CO 4319 51304 QIAamp™ DNA Mini Kit 50 prp $ 100.48 CO 4316 30210 QIAexpress™ Ni-NTA Agarose Kit 25 ml $ 176.74 CO 4158 20021 QIAEX™ II Gel Extraction Kit 150 prp $ 101.38 CO 4564 12191 QIAGEN Plasmid Giga Kit 5 prp $ 324.78 CO 4159 12162 QIAGEN Plasmid Maxi Kit 10 col $ 151.62 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 105 CO CO CO CO CO CO CO CO CO CO CO CO CO PC CO CO CO PC PC 4317 4565 4281 4161 4163 4566 4203 4567 4568 4204 4504 4255 4282 4634 4595 4256 4547 6001 6025 12163 12165 12181 12143 12123 27106 27104 28144 28706 28704 28304 28104 79654 204143 74106 74104 301305 201203 201207 QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Mega Kit QIAGEN Plasmid Midi Kit QIAGEN Plasmid Mini Kit QIAprep Spin Miniprep Kit QIAprep™ Spin Miniprep Kit QIAquick 8 PCR Purification Kit QIAquick Gel Extraction Kit QIAquick™ Gel Extraction Kit QIAquick™ Nucleotide Removal Kit (50) QIAquick™ PCR Purification Kit QIAshredder™ QuantiTect SYBR Green PCR Kit RNeasy Mini Kit RNeasy™ Mini RNA Kit SuperFect Transfection Reagent (1.2 ml) Taq DNA Polymerase Taq DNA Polymerase 25 col 100 prp 5 col 25 col 25 col 250 prp 50 rxn 400 prp 250 prp 50 prp kit 50 rxn 50 cap 200 rxn 250 prp 50 prp 3 mg/ml 250u 10x250u $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 359.77 1,327.82 187.51 177.64 122.02 253.00 58.32 538.31 329.26 70.88 70.88 70.88 48.45 296.07 768.88 174.95 171.36 50.37 1,044.95 BIO–RAD ...........................................................................................................RAD _ Technical Services: (800) 424-6723 _ Web Site: http://www.bio-rad.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: All items in the RAD catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders for RAD products must be received at the Cell Center by 2pm for 2nd day availability. _ Items ordered on Friday will be available on Tuesday _ Delivery: 2nd Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE CH 2392 1610154 30% Acrylamide/Bis 19:1 500 ml $ 39.32 CH 2393 1610156 30% Acrylamide/Bis 29:1 500 ml $ 39.32 CH 2394 1610158 30% Acrylamide/Bis 37.5:1 500 ml $ 39.32 CH 2389 1610144 40% Acrylamide/Bis 19:1 500 ml $ 45.57 CH 2390 1610146 40% Acrylamide/Bis 29:1 500 ml $ 45.57 CH 2391 1610148 40% Acrylamide/Bis 37.5:1 500 ml $ 45.57 CH 2347 1610101 Acrylamide 500 g $ 99.19 CH 2352 1610700 Ammonium Persulfate 10 g $ 10.72 CH 6058 1610786 Bio-Safe Coomasie Stain 1L $ 51.41 CH 2350 1610201 Bis-Acrylamide 50 g $ 28.60 CH 5908 1610710 BME (2-mercaptoethanol) 25ml $ 16.98 CH 5911 1610404 Bromophenol Blue 10g $ 27.70 CH 5909 1610400 Coomassie™ Brilliant Blue R-250 10g $ 16.08 KT 5941 5000112 DC Protein Assay Kit II 1 kit $ 108.13 CH 6060 1610611 Dithiothreitol (DTT) 5g $ 52.48 MS 5950 1652089 Gene Pulser® Cuvettes 0.1cm 50 pk $ 116.69 MS 4517 1652086 Gene Pulser® Cuvettes 0.2cm 50 pk $ 124.21 MS 4518 1652088 Gene Pulser® Cuvettes 0.4cm 50 pk $ 116.69 CH 2354 1610718 Glycine 1 kg $ 29.49 FM 2440 1620174 Immun-Blot PVDF 10 sht $ 55.49 MR 4515 1610324 Kaleidoscope Prestained Standards 500 µl $ 89.36 FM 2372 1620147 Nitrocellulose Membrane (.20u) 13.5x16.5cm 10 sht $ 126.00 FM 2370 1620146 Nitrocellulose Membrane (.20u) 7x8.4cm 10 sht $ 46.47 FM 2373 1620115 Nitrocellulose Membrane (.45u) 33cmx3m 1 roll $ 171.57 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 106 CH MR MR MR MR KT KT CO CO CO PG PG PG PG FM FM KT CH CH CH CH CH CH FM 2395 4633 4483 4482 4481 5937 5939 5437 5438 4522 3340 3342 3341 3343 2371 2368 5360 5910 2355 2351 2353 2397 6059 2362 1610202 161-0374 1610318 1610309 1610305 5000006 5000002 7326130 7326120 7326100 1611101 1611155 1611104 1611158 1620181 1620186 1610449 1610301 1610733 1610800 1610719 1610732 1706531 1620192 PDA Precision Plus Protein Dual Color Standard Prestained Standards Broad Prestained Standards High Prestained Standards Low Protein Assay Kit Dye Reagent Concentrate Protein Assay Kit II Quantum Maxi-Prep Kit Quantum Midi-Prep Ket Quantum™ Prep Plasmid Miniprep ReadyGel, 10% Tris-Hcl, 10 well ReadyGel, 10% Tris-Hcl, 50ul ReadyGel, 4-15% Tris-Hcl, 10 well ReadyGel, 4-15% Tris-Hcl, 50ul Sequi-Blot™ PVDF Membrane (.20u) 15x15cm Sequi-Blot™ PVDF Membrane (.20u) 7x8.4cm Silver Stain Plus Kit Sodium Dodecyl Sulfate (SDS) TBE 10X TEMED Tris TRIS/Glycine/SDS 10X Buffer Tween 20 Zeta-Probe® GT Membrane15x15cm 10 g 50 app 500 µl 500 µl 500 µl 1 kit 1 kit 10 prp 20 prp kit EA EA EA EA 10 sht 10 sht kit 100g 1L 5 ml 1 kg 1L 100 ml 15 sht $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 75.96 93.76 80.42 75.96 68.81 69.70 81.32 107.23 107.23 107.23 8.14 8.14 8.14 8.14 99.19 64.34 116.69 25.91 21.02 13.40 55.40 19.66 35.81 180.51 RAININ ................................................................................................................RAI _ Technical Services: (800) 828-2788 _ All items listed below are available on site at the Cell Center. _ SPECIAL ORDERING: Some items in the Rainin catalog may be special ordered. Orders must be received in the Cell Center before 2pm for 3rd day delivery. _ Delivery: 3rd Business Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE MS 5955 P-2 Pipetman 0.1-2.0µl 1 ea $ 266.92 MS 5956 P-10 Pipetman 0.5-10µl 1 ea $ 266.92 MS 5954 P-1000 Pipetman 100-1000µl 1 ea $ 234.45 MS 5952 P-100 Pipetman 10-100µl 1 ea $ 234.45 MS 5951 P-20 Pipetman 2-20µl 1 ea $ 234.45 MS 5953 P-200 Pipetman 50-200µl 1 ea $ 234.45 SIGMA-ALDRICH ............................................................................................................... SIG _ Technical Services: (800) 521-8956 _ Web Site: http://www.sigma-aldrich.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: All items in the Sigma catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders for Sigma products must be received at the Cell Center by 2pm for 2nd day availability. _ Items ordered on Friday will be available on Wednesday _ Delivery: 2nd Business Day PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE SM 7135 F2442-500 Fetal Bovine Serum, lot #102K8410, 110% efficiency 500 ml $ CH 5913 A8456-100mg 4-(2-Aminoethyl)Benzenesulfonyl*Fluoride Hydrochloride 100 mg $ CH 5912 A6279-10ml Aprotinin from Bovine Lung 10 ml $ University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 107 179.47 66.71 80.37 CH PC CH CH CH CH CH KT CO MB PC CH CH CH CH CH CH CH CH CH CH CH CH CH PC PC PC PC PC CH CH CH CH CH CH CH CH 2446 5411 5914 6062 2445 2375 2376 4016 4539 6067 5410 5916 5917 6063 6066 5448 5918 2403 2404 2398 2401 2402 6065 2399 6055 5477 5408 5409 5414 5922 5923 6064 5921 5919 5920 1287 1288 B2518-100mg BOVINE SERUM ALBUMIN MOLECULAR BIOLOGY GRADE 100g D7295-.5ml Deoxynucleotide Mix 1 vl D2650-5x5ml Dimethyl Sulfoxide Hybri-Max® Sterile*Filtered 25 ml D2650-100ml DMSO 100 ml D9891-25g DOXYCYCLINE HYDROCHLORIDE 25g E1510 Ethidium Bromide Aqueous Solution 10mg/ml 10 ml E4391 Ethidium Bromide Tablets 10mg 10 ea XNAT-100RXN Extract n Amp Red 100 rxn May-00 Genelute Agarose Spin Column 70 ea G4877-10000iu GONADOTROPIN FROM PREGNANT MARES' SERUM 10000iu D5062-500u Klentaq LA DNA Polymerase 500u L7533 LB Agar - EZMix® 6 x 500 ml L7658-1kg LB Broth - EZMix® 1 kg L2884-10mg Leupeptin Hemisulfate from Microbial Source 10 mg H7033-125ml Perfect Hyb Plus Hybridization Buffer 125 ml H7033-1L Perfect Hyb Plus Hybridization Buffer 1L P7626-5g Phenylmethylsulfonyl Fluoride 5g P8465-5ml Protease Inhibitor Cocktail-Bacterial 5 ml P8465-25ml Protease Inhibitor Cocktail-Bacterial 25 ml P2714 Protease Inhibitor Cocktail-General 1 btl P8340-1ml Protease Inhibitor Cocktail-Mammalian w/o Metal Chelators 1 ml P8340-5ml Protease Inhibitor Cocktail-Mammalian w/o metal chelators 5 ml P8849-5ml Protease Inhibitor Cocktail-Poly His Tagged 5 ml P8215-1ml Protease Inhibitor Cocktail-Yeast/Fungal 1 ml D4309-250u Redtaq polymerase 250 unit S4438-100rxn SYBR Green qPCR 100 rxn D1806-250u Taq DNA Polymerase 250u D4545-250u Taq DNA Polymerase w/10X Buffer 250u T4809 Taqstart Antibody 200 tst T9179 Terrific Broth (Modified)-EZMix® 6 x 500ml T9424-25ml Tri Reagent 25 ml T9424-100ml Tri Reagent 100 ml T6066-1Kg TRIZMA® Base Molecular Biology Reagent Grade 1 kg T1503-1kg TRIZMA® Base Reagent Grade 1 kg T3253-1kg TRIZMA® Hydrochloride Reagent Grade 1 kg T2444 Trizma-Hcl, 1 M Stock Solution, pH 7.6 1L T2694 Trizma-Hcl, 1 M Stock Solution, pH 8.0 1L $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ STRATAGENE ..............................................................................................................STR _ Technical Services: (800) 426-8157 _ Web Site: http://www.stratagene.com _ All items listed below are available on site at the Cell Center. _ Special Ordering: All items in the STR catalog may be special ordered, no shipping or dry ice fees. _ Conditions: All special orders for STR products must be received at the Cell Center by 2pm for next day availability. _ Items ordered on Friday will be available on Tuesday _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION SIZE CC PRICE CE 5428 230245 BL21-Codon Plus (DE3)-RIL Competent Cells ea $ 196.77 CE 5430 230255 BL21-Codon Plus (DE3)-RP Competent Cells ea $ 196.77 CE 5427 230240 BL21-Codon Plus-RIL Competent Cells ea $ 196.77 CE 5429 230250 BL21-Codon Plus-RP Competent Cells ea $ 196.77 PC 6090 600262 Herculase Enhanced DNA Polymerase 500u $ 295.62 PC 6089 600260 Herculase Enhanced DNA Polymerase 100u $ 68.03 University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 108 95.16 25.61 29.82 84.34 174.74 34.25 31.02 147.21 76.00 339.13 162.58 21.68 122.11 57.64 75.03 427.49 43.08 29.43 118.05 24.85 29.74 118.82 118.05 29.49 84.14 121.46 78.69 80.27 183.51 67.48 24.11 75.55 72.53 71.70 113.46 16.69 16.69 KT PC PC PC PC PC PC KT KT MB MB CE CE CE CE CE CE CE 4260 5400 5401 4008 5402 5403 5447 5138 5934 4551 4552 4550 4129 5418 4258 4548 4130 5517 211190 600153 600154 600380 600250 600252 600320 300385 200518 201220 201190 200152 200238 200315 200228 200314 200236 200150 pCR-Script™ Amp Cloning Kit, (3 parts, -80,-20, RT) Pfu DNA Polymerase Pfu DNA Polymerase Pfu Ultra High Fidelity PfuTurbo™ DNA Polymerase PfuTurbo™ DNA Polymerase PfuTurbo™ HotStart DNA Polymerase Prime-It® II Random Priming Kit QuickChange™ Site-Directed Mutagenesis Kit QuickHyb® Rapid Hybridization Buffer Salmon Sperm DNA SURE® 2 Supercompetent Cells SURE® Competent Cells XL 10-Gold Ultracompetent Cells XL-1 Blue Electroporation Competent Cells XL10-Gold Ultracompetent Cells XL-1-Blue Supercompetent Cells XL-2 Blue Ultra Competent Cells 1 kit 100u 500u 100 u 100u 500u 100u 30 rxn 1 kit 250 ml 10 x 1ml ea 5x0.2ml 10 x 100µl 5x80µl 5x100ul 5x0.2ml ea $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ 263.71 85.98 345.80 102.99 93.54 382.65 104.88 155.58 475.24 154.95 156.16 156.21 152.30 194.93 161.25 129.39 146.57 183.82 USB CORPORATION........................................................................USB _ Technical Services: (800) 321-9322 _ All items listed below are available on site at the Cell Center. _ Special Ordering: NO _ Items ordered on Friday will be available on Tuesday _ Delivery: Overnight PC CODE MFGR. NO. DESCRIPTION OE 5102 70092Y Alkaline Phosphatase (shrimp) CH 2113 US12387 BCIP PC 4110 70714 ddATP Termination Mix PC 4111 70716 ddCTP Termination Mix PC 4112 70718 ddGTP Termination Mix PC 4114 70720 ddTTP Termination Mix CH 2121 70468 Denhardt's Solution CH 2156 19535 NBT (Nitro Blue Tetra) PC 5241 77102 Nucleix-dATP set aq. (200uM) 0.1M OE 5147 70710 Sequenase™ dGTP labeling mix OE 5148 70754 Sequenase™ dGTP Nucleotide Kit KT 5278 70170 Sequenase™ PCR Product Seq. Kit KT 5275 70140 Sequenase™ Quick Denature Seq. Kit OE 5150 70721 Sequenase™ Reagent Kit OE 5151 70724 Sequenase™ Stop Solution OE 5239 70775z Sequenase™ v 2.0 CH 2297 US75824 Taurine SIZE 500u 100 mg 250 µl 250 µl 250 µl 250 µl 50 ml 1g 200uM 100 µl 1 kit 1 kit 1 kit 1 kit 1.2 ml 1000u 1 kg CC PRICE $ 85.48 $ 24.34 $ 25.20 $ 25.20 $ 25.20 $ 25.20 $ 37.00 $ 59.94 $ 148.99 $ 25.20 $ 119.41 $ 262.92 $ 184.10 $ 90.93 $ 17.46 $ 417.91 $ 115.06 INDEX OF STOCKED PRODUCTS MR MR MR MR MR MR 1 kb DNA Ladder 1 kb DNA Ladder 1 kb DNA Ladder 1 kb DNA Ladder - RTS - ready load 1 kb DNA Step Ladder 1 kb Plus DNA Ladder 250 µg 1000 µg 100ug 100 app 300 lns 250 µg University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 109 MR PC MR MR MR MR MR MR PC CH CH MR KT TC CH MR CH MR CH OL OL OL OL OL OL OL CH CH CH PG PG PG CH CH CH CH OL OL OL OL OL OL OL OL OL OL OL MR MR MR OL OL PG PG 10 bp DNA Step Ladder 10 mM dNTP Mix 100 bp DNA Ladder 100 bp DNA Ladder 100 bp DNA Ladder 100 bp DNA Step Ladder 100 bp Ladder - RTS - ready load 100 bp-2kb DNA Mass Ladder 10mM dNTP Mix 10X TAE 10X TBE 123 Base Pair Ladder 1st Strand cDNA Kit 2.5S Nerve Growth Factor - Mouse Natural 20% SDS Solution 200 bp DNA Step Ladder 20X SSC Buffer 25 bp DNA Ladder 2nd Strand Buffer 3' Biotinylation 1000nm 3' Biotinylation 50/200nm 3' Fluorescein 1000nm 3' Fluorescein 50/200nm 3' Phosphorylation 1000nm 3' Phosphorylation 50/200nm 3’ Spacer C3 - 200/1000nm 30% Acrylamide/Bis 19:1 30% Acrylamide/Bis 29:1 30% Acrylamide/Bis 37.5:1 3-8% NuPAGE® Tris-Acetate Gels 1.0 mm 10 well 3-8% NuPAGE® Tris-Acetate Gels 1.0 mm 12 well 3-8% NuPAGE® Tris-Acetate Gels 1.5 mm 10 well 4-(2-Aminoethyl)Benzenesulfonyl*Fluoride Hydrochloride 40% Acrylamide/Bis 19:1 40% Acrylamide/Bis 29:1 40% Acrylamide/Bis 37.5:1 5' Amine C6 TFA 5' Biotinylation 1000nm 5' Biotinylation 50/200nm 5' Fluorescein 1000nm 5' Fluorescein 50/200nm 5' or 3' Primary Amine 1000nm 5' or 3' Primary Amine 50/200nm 5' or 3' Terminal Thiol 1000nm 5' or 3' Terminal Thiol 50/200nm 5' Phosphoryation 1000nm 5' Phosphoryation 50/200nm 50 bp DNA Ladder 50 bp DNA Ladder 50 bp DNA Ladder 6-Fam, Hex, and Tet ABI dye 1000nm 6-Fam, Hex, and Tet ABI dye 50/200nm 7% NuPAGE® Tris-Acetate Gels 1.0 mm 10 well 7% NuPAGE® Tris-Acetate Gels 1.5 mm 10 well 50 µl 1 ML 50 µg 50ug 250 µl 100 µl 100 app 200 µl 100 µl 1L 1L 100 µg 30 rxn 0.1 mg 500ML 100 µl 1L 50 µg 0.5 ml ea ea ea ea ea ea ea 500 ml 500 ml 500 ml ea ea ea 100 mg 500 ml 500 ml 500 ml EA ea ea ea ea ea ea ea ea ea ea 200 µg 50 µg 100 lns ea ea ea ea University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 110 -ARE MB MB RE CH CH CH CH CH CH PC PC PC SV SV SV RE RE CH CH CH OE OE CH CH CH CH CH CH CH CH CH RE TC TC OE OE RE MS RE CH CH CH MS PC PC PC PC PC PC PC PC PC PC Aat II ABTS® Buffer ABTS® Tablets (50mg) Acc I AccuGel™ 19:1 Acetyl CoA Acetyl CoA Acrylamide Acrylamide Acrylamide Advantage cDNA PCR Kit Advantage cDNA Plolymerase Mix Advantage RT for PCR Affinity Chromatography Affinity Column Affinity Column Afl II Afl III Agar - Bacterial Select Agar - Granulated Agar - Granulated Agarase Agarase Agarose Agarose Agarose (LMP) SeaPlaque® Agarose (LMP) SeaPlaque® Agarose 1000 Agarose NuSieve® 3:1 Agarose NuSieve® GTG® Agarose SeaKem® LE Agarose SeaKem® LE Age I AIM V Medium AIM V Medium Alkaline Phosphatase (shrimp) Alkaline Phosphatase (shrimp) Alu I Aluminum Foil (18” x 50ft) AlwN I Ammonium Persulfate Ammonium Persulfate Ampicillin - 25mg/ml Amplify™ AmpliTaq Gold™ w/ buffer I - 1000u AmpliTaq Gold™ w/ buffer I - 250u AmpliTaq Gold™ w/ buffer I-12 pac (250u) AmpliTaq Gold™ w/ buffer I-5 pac (1000u) AmpliTaq Gold™ w/ buffer I-6 pac (250u) AmpliTaq Gold™ w/ buffer II - 1000u AmpliTaq Gold™ w/ buffer II - 250u AmpliTaq Gold™ w/ buffer II-12 pac (250u) AmpliTaq Gold™ w/ buffer II-5 pac (1000u) AmpliTaq Gold™ w/ buffer II-6 pac (250u) 500u 125 ml 20 tab 500u 450 ml 10 mg 10 mg 500 g 500 g 500 g 1 ea 1 ea ea per run 1 ml 5 ml 2000u 250u 500 g 500 g 500 g 100u 100u 100 g 500 g 125 g 25 g 100G 125 g 125 g 500 g 125 g 250u 500 ml 1L 1000u 500u 500u ea 500u 100 g 10 g 10 ml 1L 1000u 250u 3000u 5000u 1500u 1000u 250u 3000u 5000u 1500u University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 111 PC PC PC PC PC PC PC PC PC PC PC PC OE OE OE MB TC TC SV SV SV IM IM CO IM IM IM RE RE RE CH MS MS MS MS MS MS RE SV RE RE PC PC SV RE RE RE TC AmpliTaq® w/ buffer I - 1000u AmpliTaq® w/ buffer I - 12 paq (250u) AmpliTaq® w/ buffer I - 250u AmpliTaq® w/ buffer I - 5 paq (1000u) AmpliTaq® w/ buffer I - 6 paq (250u) AmpliTaq® w/ buffer II - 1000u AmpliTaq® w/ buffer II - 250u AmpliTaq® w/ buffer II - 5 paq (1000u) AmpliTaq® w/ buffer II - 6 paq (250u) AmpliTaq® w/ buffer II- 12 paq (250u) AmpliWax® PCR Gem 50 AmpliWax® PCR Gem 100 AMV Reverse Transcriptase AMV Reverse Transcriptase AMV Reverse Transcriptase Amylose Resin Antibiotic-Antimycotic - Pen/Strep/Fung. Antibiotic-Antimycotic - Pen/Strep/Fung. Antibody conjugation Antibody precipitation with ammonium sulfate Antibody purification (protein G) Anti-GST Antibody Anti-HA (12CA5) Anti-His Antibody Selector Kit (3 x 3 µg) Anti-Mouse Ig - HRP conjugated Anti-Rabbit Ig - HRP conjugated Anti-V5 Antibody Apa I Apa I Apa I Aprotinin from Bovine Lung ART® 10 aerosol resistant pipet tip (racked) ART® 1000E aerosol resistant pipet tip (racked) ART® 100E aerosol resistant pipet tip (racked) ART® 200 aerosol resistant pipet tip (racked) ART® 20E aerosol resistant pipet tip (racked) ART® 20P aerosol resistant pipet tip (racked) Asc I Ascites Ase I Asp 718 ATP 0.1M ATP 0.1M Autoclave Ava II Avr II Avr II Azaserine - 100X 0.6mM 0.1mg/ml 1000u 3000u 250u 5000u 1500u 1000u 250u 5000u 1500u 3000u 200 gem 200 gem 1000u 1000u 300u 100 ml 100 ml 20 ml 1 liter 0.5ml 200 µg 3x3ug 1 ml 1 ml 50 ul 5000u 2000u 5000u 10 ml 960/pk 800/pk 960/pk 960/pk 1000/pk 960/pk 500u per mouse 2000u 1000u 25 µmol 40 µmol per cycle 2000u 100u 500u 10 ml -BRE RE RE RE RE BamH I BamH I BamH I BamH I BamH I 2000u 2500u 2000u 10000u 50000u University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 112 RE RE SV CH CH CH RE RE MR CH TC RE RE RE RE RE RE PC KT KT FM FM FM CH PC PC PC CH CH CH CE CE CE CE CH NR CH SV CH MB CH MB CH RE RE RE RE RE RE RE RE RE BamH I BamH I HC Bashaw Fly Food BCIP BCIP BCIP Bcl I Bcl I BENCHMARK™ Prestained Protein Ladder Bestatin bFGF - Human Recombinant Bgl I Bgl II Bgl II Bgl II Bgl II Bgl II BigDye™ Terminator Kit BioArray High Yield RNA Transcript Labeling Kit BioArray High Yield RNA Transcript Labeling Kit Biomax™ Film MR-1 8x10in Biomax™ Film MR-1 35x43cm Biomax™ Film MS-1 8x10in Bio-Safe Coomasie Stain Biotin 11 CTP Biotin 16 UTP Biotin-16-dUTP Bis-Acrylamide Bis-Acrylamide Bis-Acrylamide BL21-Codon Plus (DE3)-RIL Competent Cells BL21-Codon Plus (DE3)-RP Competent Cells BL21-Codon Plus-RIL Competent Cells BL21-Codon Plus-RP Competent Cells Blasticidin Blocking Reagent BME (2-mercaptoethanol) Bonini Fly Food Boric Acid Bovine Serum Albumin (Nuclease free) BOVINE SERUM ALBUMIN MOLECULAR BIOLOGY GRADE Bovine Serum Albumin, Acetylated Bromophenol Blue Bsm I BspE I BspH I BssH II BstE II BstE II BstE II BstX I Bsu36 I 2500u 10000u 3ml/150mg 100 mg 100 mg 500u 3000u 2 x 250u 10 mg 10 µg 1000u 500u 400u 2000u 10000u 500u 24 rxn 10 rxn 20 rxn 50 sht 50 sht 50 sht 1L 250nm 250nm 50 nmol 25 g 50 g 25 g ea ea ea ea 50 MG 50 g 25ml 500 g 20 mg 100g 150 mg 10g 200u 1000u 500u 200u 500u 2000u 2000u 1000u 500u -CKT Cal Phos Mammalian Transfection Kit sm box University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 113 OE OE OE CH CH NR SV SV SV SV SV SV SV TC SV SV SV NR CH CH CH NR MS CH CO CO FM CH RE RE RE TC MB FM CH CH CH PC PC Calf Intestinal Alkaline Phosphatase 1000u Calf Intestinal Alkaline Phosphatase 1000u Calf Intestinal Alkaline Phosphatase 1000u Casamino Acids 500 g Casamino Acids 500 g CDP-Star Nucleic Acid Chemiluminescence Reagent (Ready to Use) 1 kit Cell Culture ATCC line Cell Culture ATCC shipping Cell culture Large scale 1 liter Cell Culture T150 Cell Culture T25 Cell Culture T75 Cell Culture–Personnel Training Cell Dissociation Buffer - Enzyme free 100 ml Cell Fusion w/ PEG EA Cell Line Flask - Set Up Ea Cell Line Flask - Weekly Charge Ea Cell Proliferation Kit (WST-1) ea Cesium Chloride 1 kg Cesium Chloride 100 g Cesium Chloride 500 g Chemiluminescent Western Blot Kit 1000cm2 Chillete personal tube cooler -12 tubes at -20C 1 ea Chloramphenicol 34 mg/ml 10 ml CHROMA SPIN-10 Columns 1 ea CHROMA SPIN-100 Columns 1 ea Chromatography Paper 3MM 46X57cm 100/pk Citric Acid 500 g Cla I 500u Cla I 1000u Cla I 500u Collagen - Rat Tail 100 mg Collagenase P 100 mg Colony/Plaque Screen™ w/orient. holes 137mm 50/bx Complete EDTA Free 20 Tabs EA Complete Mini 25 Tabs EA Coomassie™ Brilliant Blue R-250 10g CTP 0.1M 25 µmol CTP 0.1M 40 µmol -DPC PC PC PC PC KT PC PC PC PC PC PC PC RE dATP (100mM) dATP 0.1M dATP 0.1M dATP 100mM solution DATP PCR Grade DC Protein Assay Kit II dCTP (100mM) dCTP 0.1M dCTP 0.1M dCTP 100mM solution DCTP PCR Grade ddATP Termination Mix ddCTP Termination Mix Dde I 40 µmol 25 µmol 25 µmol 100 µm ea 1 kit 40 µmol 25 µmol 25 µmol 100 µm ea 250 µl 250 µl 500u University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 114 RE PC PC PC CH PC CH CH PC PC PC PC PC CE CE NR NR NR CH TC CH CH CH TC TC TC TC TC TC TC TC TC TC TC TC TC CH CH SV MR MR MR MR MR MR MR OE OE OE PC OE OE OE OE OE Dde I 200u ddGTP Termination Mix 250 µl ddNTP Set 4x0.5µmol ddTTP Termination Mix 250 µl Denhardt's Solution 50 ml Deoxynucleotide Mix 1 vl Dextran Sulfate 100 g Dextran Sulfate 100 g dGTP (100mM) 40 µmol dGTP 0.1M 25 µmol dGTP 0.1M 25 µmol dGTP 100mM solution 100 µm DGTP PCR Grade ea DH5alpha™ Library Efficiency Comp Cells 1 ml DH5alpha™ Subcloning Efficiency Comp Cells 2 ml DIG/Genius High Prime DNA Labeling and Detection Starter Kit II 1 kit Digoxigenin RNA Labeling Mix 20 rxn Digoxigenin-11-UTP 250 nmol Dimethyl Sulfoxide Hybri-Max® Sterile*Filtered 25 ml Dispase 100 ml Dithiothreitol (DTT) 5g Dithiothreitol (DTT) 5g Dithiothreitol (DTT) 5g DMEM - high glucose 25mM Hepes 500 ml DMEM - high glucose 4.5 gm/L 1L DMEM - high glucose 4.5 gm/L 500 ml DMEM - high glucose 4.5 gm/L - no sodium pyruvate 500 ml DMEM - high glucose w/o phenol red (1x) 500 ml DMEM - low glucose 500 ml DMEM H-glu w/ Glutamax 500ML DMEM Nutrient Mix F12 500 ML DMEM w/GlutaMAX 500 ML DMEM WITH L-GLUT; HI GLUCOSE; W/ NA PYRUVATE 1L DMEM WITH L-GLUT; HI GLUCOSE; W/O NA PYRUVATE 1L DMEM/F-12 - 1:1 mix w/ 15mM Hepes 500 ml DMRIE-C 1 ml DMSO 1L DMSO 100 ml DNA isolation (10-20 ml blood or 100 ml culture) DNA Marker IV (74bp - 19329bp) 50 µg DNA Marker IX (72-1353bp) 50 µg DNA Marker V (8-587bp) 50 µg DNA Marker VI (154-2176bp) 50 µg DNA Marker VIII (19bp - 1114bp) 50 µg DNA Marker X (75-12216bp) 100 µg DNA Marker XIV (100 - 1500 bp) 100 bp ladder 50 µg DNA Polymerase I 500u DNA Polymerase I from E.coli 500u DNA Polymerase I (E Coli) 1000U DNA Polymerization Mix 4x10umol DNase 1 1000u DNase I 20000u DNase I - Amplification Grade 100u DNase I - Grade II 100 mg DNase I - RNase Free 10000u University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 115 OE KT PC PC PC PC PC CH CH CH CH MB CH TC TC TC TC TC TC TC TC RE RE RE SV SV PC PC PC PC PC DNase RQ1 RNase-free DNeasy Tissue Kit dNTP Set 4x100umol dNTP Set (4x25umol) 100mM dNTP Set 0.1M dNTP's (bulk) (100mM) 4x40umoles dNTP's PCR Grade DO Supplement-His DO Supplement-Leu DO Supplement-Trp DO Supplement-Ura DOTAP (Liposomal Transfection Reagent) DOXYCYCLINE HYDROCHLORIDE DPBS (10X) with Ca++ & Mg++ DPBS (10X) without Ca++ & Mg++ DPBS (10X) without Ca++ & Mg++ DPBS (1X) with Ca++ & Mg++ DPBS (1X) with Ca++ & Mg++ DPBS (1X) with Ca++ & Mg++ DPBS (1X) without Ca++ & Mg++ DPBS (1X) without Ca++ & Mg++ Dpn I Dra I Dra III Drosophila medium Drosophila medium dTTP (100mM) dTTP 0.1M dTTP 0.1M dTTP 100mM solution DTTP PCR Grade 1000u 50 4x100µmol EA 25 µmol 1 set ea 1 ea 1 ea 1 ea 1 ea 2 ml 25g 500 ml 4L 500 ml 1L 1L 1L 1L 1L 1,000u 2000u 1000u 100 vials 25 bottles 40 µmol 25 µmol 25 µmol 100 µm ea -EOE RE SV NR NR NR NR NR RE RE RE RE RE RE RE RE RE RE RE CH CH E. Coli DNA Ligase Eag I (Xma III) EBV stock ECL™ Plus Western Blotting Detection Reagents ECL™ Western Blot Analysis Starter System ECL™ Western Blotting Detection Reagents ECL™ Western Blotting Detection Reagents ECL™ Western Blotting Detection Reagents EcoN I EcoR I EcoR I EcoR I EcoR I EcoR I EcoR I EcoR I HC EcoR V EcoR V EcoR V EDTA 0.5M (4 x 100ml) EDTA Disodium Salt Dihydrate 100u 500u per tube 1 ea 1 kit 1000cm2 4000cm2 2000cm2 1000u 5000u 5000u 20000u 10000u 50000u 5000u 10000u 2000u 2000u 2000u 400 ml 500 g University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 116 CO CE CE MS MS MS FM FM OE CO SV SV TC TC SV SV SV SV SV SV SV SV SV TC CH CH CH OE OE PC PC PC PC PC PC MB KT MB Effectene Transfection Reagent (1 ml) 1 ml ElectroMAX DH10B Efficiency Comp Cells 5x.1ml ElectroMAX DH5alpha-E Efficiency Comp Cells 5x.1ml Electroporation Cuvettes 0.1cm Electroporation Cuvettes 0.2cm Electroporation Cuvettes 0.4cm Elite II Slide Film 100 - 24 Exposures 1 roll Elite II Slide Film 100 - 36 Exposures 1 roll Elongase Enzyme Mix 100 rxn EndoFree Plasmid Maxi Kit (10) kit Endotoxin test (1-2 tests per test) Endotoxin test (3 or more tests per test) Epidermal Growth Factor 100 ug Epidermal Growth Factor - Culture Grade 0.1 mg ES Cell Culture & Karyotyping per culture ES Cell Culture for blastocystin injection ES Cell Culture T150 flask ES Cell Culture T25 flask ES Cell Culture T75 flask ES Cell Electroporation + Selection fewer than 20 clones ES Cell EP + Selection, at least 2 80% male chimera ES Cell EP+Selection, 150-200 +ve clones, no Recombinants ES Cell EP+Selection, at least 2 Recombinant clones Essential Amino Acids - 50X MEM supplemt 100 ml Ethidium Bromide (ETBR) 10mg/ml 10 ml Ethidium Bromide Aqueous Solution 10mg/ml 10 ml Ethidium Bromide Tablets 10mg 10 ea Exonuclease I 2500u Exonuclease III 5000u Expand™ High Fidelity PCR System 100u Expand™ High Fidelity PCR System 500u Expand™ High Fidelity PCR System 10x250u Expand™ Long Template PCR System 100u Expand™ Long Template PCR System 10x250u Expand™ Long Template PCR System 500u Express Hyb 250 ml Extract n Amp Red 100 rxn ExtremeGENE (Transfection Reagent) .4 ml -FTC PC PC CO SM SM SM SM SM SM SM TC TC TC F12 Nutrient Mix, Kaighns Mod FastStart Taq DNA Polymerase FastStart Taq DNA Polymerase FastTrack® 2.0 Kit Fetal Bovine Serum Fetal Bovine Serum, ES Qualified, Lot # A38507V Fetal Bovine Serum, Lot # 1185797 Fetal Bovine Serum, Lot # SFB30-1491 Fetal Bovine Serum, lot #102K8410, 110% efficiency Fetal Bovine Serum, lot #FBU2110, 145% efficiency Fetal Bovine Serum, Lot# ANG19178 Fibronectin - Human Fibronectin - Human Fibronectin, human plasma 500 ML 100u 500u 6 rxn 500 ML 500 ML 500ml 500ml 500 ml 500ML 500ml 1 mg 1 mg 5mg University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 117 CH CH KT MB CH CH SV TC MB MB FM FM TC SV SV SV SV Ficoll-Paque® Plus solution Ficoll-Paque® solution First Strand cDNA Synthesis Kit fmol® DNA Sequencing System Formamide Formamide Freezing Cells Freezing Mix - 95% Calf Serum 5% DMSO FuGENE 6 (Nonliposomal Transfection Reagent) FuGENE 6 (Nonliposomal Transfection Reagent) Fuji RX Film NIF 14x17in Fuji RX Film NIF 8x10in Fungizone Fusion - Cell Culture Work Fusion - Cell Culture Work Fusion without Immunization Fusion without Immunization 6x500ml 6x100ml 1 kit 1 kit 500 ml 500 ml per vial 100 ml .4 ml 1 ml 100 sht 100 sht 20ml 20 clones 40 clones 20 clones 40 clones G418 G418 G418 Sulfate (50mg/ml) GC Rich Polymerase Gene Pulser® Cuvettes 0.1cm Gene Pulser® Cuvettes 0.2cm Gene Pulser® Cuvettes 0.4cm GeneAmp® dNTP blend 10mM GeneAmp® dNTP blend 10mM-12 paq GeneAmp® dNTP blend 10mM-6 paq GeneAmp® Core Kit GeneAmp® dNTP Set -10mM (4x3.2umol) GeneAmp® EZ rTth RNA PCR Kit GeneAmp® RNA PCR Core Kit GeneAmp® XL PCR Kit GeneClean Turbo GeneClean® DNA Purification Spin Kit GeneClean® DNA Purification Spin Kit GeneClean® II DNA Purification Kit GeneClean® III DNA Purification Kit Genelute Agarose Spin Column Genescreen Plus® Hybridization Mem. 20x20cm Genescreen Plus® Hybridization Mem. 30x45cm Genescreen™ Hybridization Mem. 20x20cm Geneticin (G418)® Geneticin (G418)® Geneticin® - (50mg/ml) GENIUS™ 4 RNA Labeling Kit Gentamicin Glutamax I, 100X Glutathione Sepharose Glycerol Glycine Glycine Glycine 1g 5g 20 ml 50 rxn 50 pk 50 pk 50 pk 1 ml 12 ml 6 ml 100 rxn 1 set 100 rxn 100 rxn 100 rxn 50 prp 50 prp 100 prp 300 prp 600 prp 70 ea 10/bx 5 sht 10/bx 5g 1g 20 ml 2x10rxn 10 ml 100ML 10 ml 500 ml 1 kg 1 kg 500 g -GTC TC TC PC MS MS MS PC PC PC PC PC PC PC PC CO CO CO CO CO CO FM FM FM TC TC TC NR TC TC CH CH CH CH CH University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 118 CH MB MB TC KT KT PC CH CH CH Glycine Glycogen GONADOTROPIN FROM PREGNANT MARES' SERUM Grace's/Hink's supp - yeastolate+lactalbumin GST Detection Module GST Purifications Bulk Module GTP 0.1M Guanidine HCL Guanidine Isothiocyanate Guanidine Thiocyanate 1 kg 20 mg 10000iu 500 ml 50 rxn 5 prps 25 µmol 500 g 250 g 500 g Hae II Ham's F-10 Ham's F-12 Ham's F-12 Ham's F-12 Hank's BSS (10X) Hank's BSS (1X) w/o Ca++ or Mg++ or phenol Hank's BSS 1X - w/20mM hepes/no Ca&Mg HB101 High Efficiency Competent Cells HBSS HBSS w/ ca and mg HBSS w/o ca and mg HEPES HEPES Buffer Solution (1M) Herculase Enhanced DNA Polymerase Herculase Enhanced DNA Polymerase Hha I Hha I Hha I High Prime DNA Labeling Kit High Prime DNA Labeling mix for dCTP High Pure Maxi Kit High Pure PCR Template Prep High Pure Plasmid Isolation Kit High Pure Plasmid Isolation Kit Hinc II Hind III Hind III Hind III Hind III Hind III Hind III HC Hinf I Hispeed Midi Kit (25) 45 minutes/200ug Hispeed Plasmid Maxi Kit Hispeed Plasmid Maxi Kit Hollow Fiber cartridge setup Hollow Fiber cartridge setup Hollow Fiber run (5% FBS) Hollow Fiber Run (BD Serum Free) Horse Serum Hot Star Taq (1000) with Q solution 2000u 500 ml 500 ml 1L 500 ml 500 ml 1L 1L 1 ml 500 ML 1L 1L 100 g 100 ml 500u 100u 1500u 2000u 1000u 50 rxn 50 rxn 10rxn EA 50 col 250 col 1000u 5000u 10000u 5000u 10000u 5000u 10000u 5000u 45min/200ug 10 prep 25 prep medium large per week per week 500 ml (4x250u) -HRE TC TC TC TC TC TC TC CE TC TC TC CH CH PC PC RE RE RE KT KT KT KT CO CO RE RE RE RE RE RE RE RE CO KT KT SV SV SV SV SM PC University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 119 PC PC PC PC RE RE RE MB FM FM FM FM FM FM FM FM FM FM FM FM FM FM FM TC TC TC TC FM FM FM FM FM TC TC Hot Star Taq (250) with Q solution HotMaster Taq w/ buffer HotMaster Taq w/ buffer HotMaster Taq w/ buffer Hpa I Hpa I Hpa II Human Cot-1® DNA Hybond™ ECL™ 20x20cm Hybond™ ECL™ 30cmx3m Hybond™ Nylon 137mm Hybond™ Nylon 20cmx3m Hybond™ Nylon 22x22cm Hybond™ Nylon 30cmx3m Hybond™ Nylon 82mm Hybond™ Nylon 87mm Hybond™-C extra sup nitrocellulose 30cmx3m Hybond™-N+ charged nylon 137mm Hybond™-N+ charged nylon 20cmx3m Hybond™-N+ charged nylon 20x20cm Hybond™-N+ charged nylon 30cmx3m Hybond™-N+ charged nylon 87mm Hybond™-PVDF 20x20cm Hybridoma Growth Supp - low endotoxin 10X Hybridoma Growth Supplement - High Endotoxin 10–50X Hygromycin B (50mg/ml) Hygromycin B (50mg/ml) Hypercassette™ (beige) 35x43cm Hyperfilm MP Enveloped, 8x10in Hyperfilm™ ECL™ 8x10in Hyperfilm™ ECL™ 8x10in Hyperfilm™ MP 8x10in Hypoxanthine - 100X 10mM 1.36mg/ml Hypoxanthine/Thymidine - 100X 10mM/1.6mM 250u 100u 250u 1000u 500u 100u 2000u 500 µg 10 sht 1 roll 50/bx 1 roll 10/bx 1 roll 50/bx 50/bx 1 roll 50/bx 1 roll 10 sht 1 roll 50/bx 10 sht 25 ml 25 ml 20 ml 20 ml 1 ea 50 sheets 25 sht 75 sht 75 sht 10 ml 50 ml -I- -J- -KTC TC TC TC CH CH CH FM FM FM FM SV NR TC CH CH CH SM TC IL-2 Human Recombinant IL-2 Mouse Recombinant IL-4 - Human Recombinant IMDM w/GlutaMAX imMedia™ Amp Blue imMedia™ Kan Blue imMedia™ Zeo Agar Immobilon-NY+ 137mm DISC(Colony Plaque) Immobilon™ P 20x20cm (Western Blot) Immobilon™ P 26.5cmx3.75m (Western Blot) Immun-Blot PVDF Immunization In Situ Cell Death Detection Kit - Fluorescein Interleukin 2 - 10000u Human recombinant IPTG IPTG IPTG Iron Enriched Calf Serum — Lot AKH12367 Iscove's Medium 10000u 10000u/1ml 5 µg 500 ML ea ea ea 50/pk 10/pk 1 roll 10 sht per mouse 50 rxn 1 ml 1g 1g 10 g 500 ml 500 ml University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 120 TC TC TC CE MR TC TC TC OE OE OE OE OE PC RE RE RE RE Iscove's Medium ITS Premix - Culture Supplement ITS+ Premix - Culture Supplement JM109 High Efficiency Competent Cells Kaleidoscope Prestained Standards Kanamycin - 100X - 10mg/ml Kanamycin/Neomycin - 100X - 20 K/5 N mg/ml Kennett's HY - 90% DMEM hg/10% NCTC 135 Klenow - Exo Minus Klenow - Labeling grade Klenow - Sequencing Grade Klenow - Sequencing Grade Klenow Fragment Klentaq LA DNA Polymerase Kpn I Kpn I Kpn I Kpn I HC 500 ml 5 ml 20 ml 1 ml 500 µl 10 ml 25 ml 1L 100u 100u 100u 200u 150u 500u 5000u 3000u 2500u 2000u Lambda DNA Lambda DNA/HindIII Fragment Lambda DNA/HindIII Fragment Lambda DNA/HindIII Fragment Lambda DNA/HindIII Fragment - RTS - ready load Lambda Quick! Phage DNA Isolation Spin Kit Laminin - Mouse LB + Ampicillin (50ug/ml) Agar Plates LB + Kanamycin (50ug/ml) Agar Plates LB + Tetracycline (20ug/ml) Agar Plates LB Agar LB Agar - EZMix® LB Agar - Miller LB Agar - Miller LB Agar Plates LB BROTH LB Broth - EZMix® LB Broth - Lennox LB Broth - Lennox LB Broth - Miller LB Broth - Miller LB Medium Capsules LC FastStart DNA Mstr Sybrgreen LC FastStart DNA Mstr Sybrgreen Leupeptin Leupeptin Hemisulfate from Microbial Source L-Glutamine - 100X - 200mM - 29.23mg/ml L-Glutamine - 200mM L-Glutamine - 200mM LipofectAMINE 2000 LipofectAMINE 2000 Lipofectamine™ Lipofectamine™ Lipofectamine™ Lipofectin® Reagent 250 µg 500 µg 150 µg 100µg 100 app 25 prp 1 mg 10 pl 10 pl 10 pl 500 g 6 x 500 ml 500 g 500 g 10 pl 1L 1 kg 500 g 500 g 500 g 500 g 1 kg EA EA 25 mg 10 mg 100 ml 100 ml 100 ml 1.5ml 0.75ml 1 ml 4 x 1 ml .5 ml 1 ml -LMR MR MR MR MR CO TC CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH PC PC CH CH TC TC TC MB MB MB MB MB MB University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 121 SV NR NR NR NR NR SV SV Liquid N2 storage (per vial/quarter) Luciferase Assay System Luciferase Assay System (10 pk) quote#5075 Luciferase Assay System–10 pack Lumi Light Lumi Light + Lymphocyte EBV Transformation Lymphocyte isolation 100 assays 1000 assays 1000 assays 1 kit 1 kit -MMB RE RE MR MR FM FM CH OE OE OE MR PC PC PC PC TC TC TC TC TC CE CE CE RE RE TC TC TC TC TC TC TC TC RE CO CO CO CO CO MS MS CH CH m7GpppG 5'G RNA Cap Structure Mae I Mae II MagicMark Western Protein Standard MagicMark XP Magna 137mm Notched Magnagraph 137mm Magnesium Chloride Maloney-MLV Reverse Transcriptase Maloney-MLV Reverse Transcriptase Maloney-MLV Reverse Transcripts Mark12™ Wide Range Molecular Weight Standard MasterMix MasterTaq Kit MasterTaq Kit MasterTaq Kit Matrigel® Growth Factor Reduced Matrigel® Matrix Matrigel® Phenol Free & Growth Fact Reduced Matrigel® Phenol Red Free Matrisperse® Cell Recovery Solution Max Efficiency DH5@T1 Competent Cells MAX Efficiency DH5alpha Efficiency Comp Cells Max Efficiency STBL2 Mbo I Mbo I McCoy's 5A Medium 199 MEM - w/ Earles' salts & L-Glut MEM - w/ Earles' salts & L-glut MEM - w/ Earles' salts & L-Glut MEM alpha - w/ nucleosides MEM alpha - w/o nucleosides MEM alpha - w/o nucleosides Mfe I Micro-FastTrack™ Kit MicroSpin G-25 Columns MicroSpin G-50 Columns MicroSpin S-200 HR Columns MicroSpin S-300 HR Columns Mini-Centrifuge Mini-Centrifuge 6 x 1.5 ml Minimal SD Agar Base Minimal SD Base 25u 50u ea 250ul 250ul 50/pk 50/pk 500 g 200u 40000u 10000u 1.0 ml 152u 100u 250u 500u 10 ml 10 ml 10 ml 10 ml 100 ml 1 ml (5 x 0.2ml) 5x.1ml 1 ML 500u 200u 500 ml 500 ml 1L 1L 1L 1L 1L 500 ml 500u 20 rxn 50 col 50 col 50 col 50 col ea 1 ea 1 ea 1 ea University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 122 RE RE RE RE MR OE IM SV SV SV SV Mlu I Mlu I Mnl I Msc I MultiMark® Multi-Colored Standard Mung Bean Nuclease myc 1 - 9E10 antibody Mycoplasma Enzymatic test, MycoAlert Mycoplasma Enzymatic test, MycoAlert Mycoplasma test PCR (1-2 samples per sample) Mycoplasma test PCR (3 or more samples per sample) 1000u 1000u 250u 100u 500 µl 3000u 0.5 mg 1-5 samples > 5 samples N2 Supplement Nae I Nar I NBT (Nitro Blue Tetra) NBT (Nitro Blue Tetra) Nco I Nco I Nco I Nde I Nde I Neural Supplement - B27 (50X) Neuralbasal Media Nhe I Nhe I Nhe I Nick translation Kit NICK® Columns Ni-NTA Superflow Nitrobind 7x8.4 Nitrobind Pure 137mm Nitrobind Pure 20cmx3m Nitrobind Pure 30cmx3m Nitrocellulose 82mm Colony & Plaque Nitrocellulose 137mm Colony & Plaque Nitrocellulose Membrane (.20u) 13.5x16.5cm Nitrocellulose Membrane (.20u) 7x8.4cm Nitrocellulose Membrane (.45u) 33cmx3m Nitrocellulose Membrane (0.2 µm) Filter Paper Sandwich Nitrocellulose Membrane (0.45 µm) Filter Paper Sandwich Nitropure Supp 82mm Nitropure Supp 30cmx3m Non-essential amino acids - 100X Not I Not I Not I Not I Not I NP Transfer Buffer (20X) Nru I Nru I Nsi I 5 ml 500u 200u 3 ml 1g 200u 1000u 200u 4000u 500u 10 ml 500 ml 1000u 5000u 250u 50 rxn 50 col 25 ml 10/pk 50/pk 1 roll 1 roll 50/pk 50/pk 10 sht 10 sht 1 roll 20/pk 20/pk 50/pk 1 roll 100 ml 200u 200u 500u 2500u 200u 1L 1000u 200u 1000u -NMB RE RE CH CH RE RE RE RE RE TC TC RE RE RE KT CO CO FM FM FM FM FM FM FM FM FM FM FM FM FM TC RE RE RE RE RE CH RE RE RE University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 123 PC PC PG PG PG PG PG PG PG PG PG PG CH CH CH CH CH CH CH TC NTP Set 0.1M Nucleix-dATP set aq. (200uM) 0.1M NuPAGE 12% Bis-Tris Gels 1.0mm 12 well NuPAGE 4-12% Bis-Tris Gels 1.5mm 10 well NuPAGE® 10% Bis-Tris Gels 1.0 mm 10 well NuPAGE® 10% Bis-Tris Gels 1.0 mm 12 well NuPAGE® 10% Bis-Tris Gels 1.0 mm 15 well NuPAGE® 12% Bis-Tris Gels 1.0 mm 10 well NuPAGE® 12% Bis-Tris Gels 1.0 mm 15 well NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 10 well NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 12 well NuPAGE® 4-12% Bis-Tris Gels 1.0 mm 15 well NuPAGE® Antioxidant NuPAGE® LDS Sample Buffer (4X) NuPAGE® MES SDS Running Buffer (20X) NuPAGE® MOPS SDS Running Buffer (20X) NuPAGE® Sample Reducing Agent (10X) NuPAGE® Transfer Buffer (20X) NuPAGE® Tris-Acetate SDS Running Buffer (20X) Nutridoma® SP 4x25µmol 200uM ea EA ea ea ea ea ea ea ea ea 15ml 10ml 500ml 500ml 250ul 125ml 500ml 10 ml -O- -PMB MS MS MB MB CE CE CE CE CE TC TC TC MS MS MS MS MS RE MB KT PC PC PC PC KT CH CH TC TC TC TC Oligo (dT) 12-18 Primer Oligo CD - $1,000 Oligo CD - $500 Oligo(dT) 15 primer Oligofectamine Reagent One Shot MACH 1 Comp eColi (LTI Shelf) One Shot® MAX Efficiency® DH5a®-T1® OneShot™ INVaF' Competent Cells OneShot™ TOP10 Competent Cells OneShot™ TOP10F' Competent Cells OPI 100X - 15mg O - 5 mg P - 20u I/ml Opti-MEM® - reduced serum hybrid. medium Origen® Hybridoma Cloning Factor - 10-50X P-100 Filter Pipet Tips P-1000 Filter Pipet Tips P2/P10 Eatra Long Filter Pipet Tip P-20 Filter Pipet Tips P-200 Filter Pipet Tips Pac I pBR322/Msp I Digest pcDNA3.1© Directional TOPO® Expression Kit (Top 10 Cells) PCR Cleanup Kit PCR Markers PCR Nucleotide Mix PCR Optimization Kit (for optimal pH & Mg) pCR-Script™ Amp Cloning Kit, (3 parts, -80,-20, RT) PDA Pefabloc® SC PEG(50%+5%DMSO) - hi efficiency/fusion Pen/Strep - 100X -10000u/ml P - 10mg/ml S Pen/Strep - 100X -10000u/ml P - 10mg/ml S Pen/Strep - 100X -10mcg/ml P - 10mg/ml S 25 UG EA EA 20 µg 1 ML 20 rxn 20 x 50ul 21x50µl 21x50µl 21x50µl 50 ml 500 ml 50 ml EA EA EA EA EA 250u 50 µg 20 rxn 100 rxn 250 µl 100 rxn 1 kit 1 kit 10 g 100 mg 2 ml 100 ml 100 ml 100 ml University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 124 CH CH CH CH CH CH TC TC PC PC PC PC PC PC MB MB MB CH CH CH CH CH CH CH CH CH MR MR MR MR MR MS MS MS MS MS MS MS MS MS MS MS MS PC PC PC PC PC PC PC PC MB RE FM FM Pepstatin (was #600160) Peptone Peptone Percoll® Perfect Hyb Plus Hybridization Buffer Perfect Hyb Plus Hybridization Buffer PFHM II - Protein Free Hybridoma Medium PFHM II - Protein Free Hybridoma Medium Pfu DNA Polymerase Pfu DNA Polymerase Pfu Ultra High Fidelity PfuTurbo™ DNA Polymerase PfuTurbo™ DNA Polymerase PfuTurbo™ HotStart DNA Polymerase pGEM®-T Easy I pGEM®-T Easy II pGEM®-T Vector I Phenol Phenol Saturated (Buffer) Phenol Saturated (Tris) Phenol (was 100300) Phenol Saturated pH 6.6 Phenol Saturated pH 6.6 Phenol/Chloroform/Isoamyl Alcohol Phenol/Chloroform/Isoamyl Alcohol Phenylmethylsulfonyl Fluoride Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment Phi-X174 DNA/Hae III Fragment - RTS - ready load Pipetman 0.1-2.0µl Pipetman 0.5-10µl Pipetman 100-1000µl Pipetman 10-100µl Pipetman 2-20µl Pipetman 50-200µl Pipet-mate Digital Pipette w/ tip ejector 10µl Pipet-mate Digital Pipette w/ tip ejector 100µl Pipet-mate Digital Pipette w/ tip ejector 1000µl Pipet-mate Digital Pipette w/ tip ejector 20µl Pipet-mate Digital Pipette w/ tip ejector 200µl Pipet-mate Digital Pipette w/ tip ejector 5000µl Platinum Pfx DNA Polymerase Platinum Pfx DNA Polymerase Platinum Pfx DNA Polymerase Platinum Supermix Hi Fi Platinum Taq DNA Polymerase - 100u Platinum Taq DNA Polymerase - 250u Platinum Taq DNA Polymerase - 500u Platinum Taq High Fidelity Plus Reagent Pme I Polaroid Film Type 57 Polaroid Film Type 667 10 mg 500 g 500 g 1L 125 ml 1L 1L 500 ml 100u 500u 100 u 100u 500u 100u 1 kit 1 kit 20u 500 g 100 ml 100 ml 500 g 100 ml 400 ml 100 ml 400 ml 5g 40 µg 250u 50u 50 µg 100 app 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 1 ea 100 u 250 u 500 u 100 rxns 100u 250u 500u 100u .85 ml 500u pk pk University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 125 MB CO FM CH MR MR MR MR MR KT MB MB MB MB MB MB MB MB MB MB CO CH CH CH CH CH CH CH CH MB IM KT KT IM CH CH CH CH CH CH CH CH CH RE RE RE KT FM RE RE RE RE RE RE Poly d(I-C) PolyATtract® System 1000 RNA Isolation System Polyscreen® PVDF 30cmx3m Potassium Chloride Precision Plus Protein Dual Color Standard Prestained Protein Molecular Weight Marker Prestained Standards Broad Prestained Standards High Prestained Standards Low Prime-It® II Random Priming Kit Primer gt10 forward Primer gt10 reverse Primer gt11 forward Primer gt11 reverse Primer M13/pUC Forward 23mer Primer M13/pUC Reverse 23mer Primer Sp6 20mer Primer T3 20mer Primer T7 20mer Primer T7 Term ProbeQuant™ G-50 Micro Columns Protease Inhibitor Cocktail Tablets (Complete) Protease Inhibitor Cocktail-Bacterial Protease Inhibitor Cocktail-Bacterial Protease Inhibitor Cocktail-General Protease Inhibitor Cocktail-Mammalian w/o Metal Chelators Protease Inhibitor Cocktail-Mammalian w/o metal chelators Protease Inhibitor Cocktail-Poly His Tagged Protease Inhibitor Cocktail-Yeast/Fungal Protector RNase Inhibitor Protein A Agarose Protein Assay Kit Dye Reagent Concentrate Protein Assay Kit II Protein G Agarose Proteinase K Proteinase K Proteinase K Proteinase K Proteinase K Proteinase K Solution (20 mg/ml) ProtoGel® (30% Acrylamide/0.8% Bis) ProtoGel® Buffer ProtoGel® Stacking Buffer Pst I Pst I Pst I PureTaq RTG PCR Beads PVDF Membrane/Filter Paper Sandwich Pvu I Pvu I Pvu I Pvu II Pvu II Pvu II 10 Å260u 1 kit 1 roll 500 g 50 app 175 lns 500 µl 500 µl 500 µl 30 rxn 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 5 µg 50 col 20 tabs 5 ml 25 ml 1 btl 1 ml 5 ml 5 ml 1 ml EA 5 ml 1 kit 1 kit 5 ml 100 mg 25 ml 500 mg 100 mg 500 mg 5 ml 1L 450 ml 200 ml 3000u 10000u 3000u 0.5ML 20/pk 150u 100u 100u 1000u 5000u 1000u University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 126 -QCO CO CO CO CO CO CO CO CO CO CO CO CO CO CO CO CO CO CO PC CO CO CO CO CO CO CO CO KT MB QIAamp™ Blood Kit QIAamp™ DNA Mini Kit QIAexpress™ Ni-NTA Agarose Kit QIAEX™ II Gel Extraction Kit QIAGEN Plasmid Giga Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Maxi Kit QIAGEN Plasmid Mega Kit QIAGEN Plasmid Midi Kit QIAGEN Plasmid Mini Kit QIAprep Spin Miniprep Kit QIAprep™ Spin Miniprep Kit QIAquick 8 PCR Purification Kit QIAquick Gel Extraction Kit QIAquick™ Gel Extraction Kit QIAquick™ Nucleotide Removal Kit (50) QIAquick™ PCR Purification Kit QIAshredder™ QuantiTect SYBR Green PCR Kit Quantum Maxi-Prep Kit Quantum Midi-Prep Ket Quantum™ Prep Plasmid Miniprep Quick Spin™ G25 DNA Quick Spin™ G25 RNA Quick Spin™ G50 DNA Quick Spin™ G50 DNA Quick Spin™ G50 RNA QuickChange™ Site-Directed Mutagenesis Kit QuickHyb® Rapid Hybridization Buffer 50 prp 50 prp 25 ml 150 prp 5 prp 10 col 25 col 100 prp 5 col 25 col 25 col 250 prp 50 rxn 400 prp 250 prp 50 prp kit 50 rxn 50 cap 200 rxn 10 prp 20 prp kit 20 col 20 col 20 col 50 col 20 col 1 kit 250 ml Rabbit Reticulocyte Lysate Rad Prime Labeling Kit Rainbow Marker™ (14300-250000) Rainbow Marker™ (2350-46000) Random Primed DNA Labeling Kit Random Primers (N)6 Random Primers (N)6 Rapid Gel Extraction Kit Rapid PCR Purification Kit Rapid Plasmid Mini Prep RapidHyb Hybridization™ Buffer Ready Load 1KB Plus DNA Ladder ReadyGel, 10% Tris-Hcl, 10 well ReadyGel, 10% Tris-Hcl, 50ul ReadyGel, 4-15% Tris-Hcl, 10 well ReadyGel, 4-15% Tris-Hcl, 50ul Ready-To-Go DNA Labelling Beads (-dCTP) Rediprime™ kit for dCTP Redtaq polymerase Renaissance™ Nucleic Acid Chemilum Rgt Kit Renaissance™ Western Blot Chemilum Rgt Renaissance™ Western Blot Chemilum Rgt 1 ml 30 rxn 250 µl 250 µl 50 rxn 50 Å260u 50 Å260u 50rxn 50rxn 50rxn 125 ml 100 rxn EA EA EA EA 1 kit 30 rxn 250 unit 1 kit 1000cm2 2500cm2 -RMB KT MR MR KT MB MB KT KT KT MB MR PG PG PG PG KT OE PC NR NR NR University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 127 NR OE OE MR MB OE OE MB OE OE MB MB MB MB CO CO TC TC TC TC TC TC TC CO RE PC PC PC Renaissance™ Western Blot Chemilum Rgt Ribonuclease H Ribonuclease H RNA Ladder 0.24-9.5 kb RNAguard Ribonuclease Inhibitor RNase - DNase free Rnase A RNase AWAY RNase H RNase T1 RNasin® Recombinant Ribonuclease Inhibitor RNasin® Recombinant Ribonuclease Inhibitor RNasin® Ribonuclease Inhibitor RNasin® Ribonuclease Inhibitor RNeasy Mini Kit RNeasy™ Mini RNA Kit RPMI 1640 RPMI 1640 RPMI 1640 RPMI 1640 - no phenol RPMI 1640 - w/ 25mM Hepes RPMI 1640 - w/ 25mM Hepes RPMI w/GlutaMAX RPM™ Rapid Pure Plasmid Miniprep Kit Rsa I rTth DNA Polymerase & EZ Buffer Pack rTth DNA Polymerase with Buffer rTth DNA Polymerase-XL w/XL Buffer Pack 5000cm2 30u 50u 75 µg 5000u 500 µg 100 mg 8.4 oz 250u 100000u 10000u 2500u 10000u 2500u 250 prp 50 prp 1L 1L 1L 500 ml 500 ml 500 ml 500 ML 120 prp 1000u 100 rxn 100 rxn 100 rxn -STC OE RE RE RE RE RE RE RE RE RE MB RE RE RE RE RE CH MR MR CH CH CH CH S.O.C. Transformation Media - 10x10 ml S1 Nuclease Sac I (Sst I) Sac I (Sst I) Sac I (Sst I) Sac II Sac II Sal I Sal I Sal I Sal I Salmon Sperm DNA Sap I Sau 3A I Sau 3A I Sca I Sca I SDS 10% (4x100ml) SeeBlue® Plus2 Pre-Stained Standard SeeBlue® Pre-Stained Standard SequaGel® Buffer SequaGel® Concentrate (20% Acryl.) 19:1 SequaGel® Diluent SequaGel® Sequencing System 100 ml 10000u 1000u 2000u 1000u 2000u 500u 500u 2000u 10000u 2000u 10 x 1ml 50u 200u 100u 500u 1000u 400 ml 500 µl 500 µl 200 ml 1L 1L 1 kit University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 128 CH CH OE OE KT KT OE OE OE FM FM TC TC RE RE NR KT MR RE RE RE RE RE RE TC CH CH CH TC TC OE OE OE OE RE RE RE SV SV SV RE RE RE RE CH CH RE RE CH RE FM FM FM FM FM SequaGel®-6 6% Monomer Sequencing Soln (includes EC-841) 1 L SequaGel®-8 8% Monomer Sequencing Soln (includes EC-841) 1 L Sequenase™ dGTP labeling mix 100 µl Sequenase™ dGTP Nucleotide Kit 1 kit Sequenase™ PCR Product Seq. Kit 1 kit Sequenase™ Quick Denature Seq. Kit 1 kit Sequenase™ Reagent Kit 1 kit Sequenase™ Stop Solution 1.2 ml Sequenase™ v 2.0 1000u Sequi-Blot™ PVDF Membrane (.20u) 15x15cm 10 sht Sequi-Blot™ PVDF Membrane (.20u) 7x8.4cm 10 sht SF900 II - Serum-free insect medium 500 ml SF900 II - Serum-free insect medium 1L Sfi I 250u Sfi I 2000u Sheep Anti-digoxigenin-AP 150u Silver Stain Plus Kit kit SimplyBlue™ SafeStain 1L Sma I 1000u Sma I 1000u Sma I 2000u Sma I 1000u SnaB I 500u SnaB I 100u Sodium Bicarbonate 100 ml Sodium Dodecyl Sulfate 100 g Sodium Dodecyl Sulfate (SDS) 500 g Sodium Dodecyl Sulfate (SDS) 100g Sodium Pyruvate - 100X 11g/l 100mM 100 ml Sodium Pyruvate, 100mM solution 100 ml SP6 RNA Polymerase 3000u Sp6 RNA Polymerase 500u Sp6 RNA Polymerase 1000u SP6 RNA Polymerase (25u/µl) 10000u Spe I 200u Spe I 2500u Spe I 200u Special Media (10L or more powder provided) 1 liter Special Media (10L or more) 1 liter Special Media (less than 10L) 1 liter Sph I 200u Sph I 200u Sph I 250u Sph I 200u SSC 20X 4L SSC 20X 4L Ssp I 1000u Ssp I 500u SSPE 20X 4L Sst I 2000u Stericup Filtr. & Stor.-PES 250ml .22µ 12/pk Stericup Filtr. & Stor.-PES 500ml .22µ 12/pk Stericup Filtr. & Stor.-PES 1000ml .22µ 12/pk Stericup Filtr. & Stor.-PES 150ml .22µ 12/pk Steriflip-GP Filter Unit 50µl 25/pk University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 129 SV FM FM FM FM IM RE RE RE RE CH CH CO OE KT OE OE PC OE PC OE CE CE RE PC PC PC FM FM FM FM Sterile Hood Usage Steritop-GP Filtr 1000 ML 33 mm neck 0.22µm Steritop-GP Filtr 1000 ML 45 mm neck 0.22µm Steritop-GP Filtr 500 ML 33 mm neck 0.22µm Steritop-GP Filtr 500 ML 45 mm neck 0.22µm Streptavidin Red 670™ Stu I Stu I Sty I Sty I Sucrose Sucrose SuperFect Transfection Reagent (1.2 ml) Superscript Custom Choice Kit Superscript First-Strand Synthesis System for RT-PCR Superscript III Reverse Transcriptase SuperScript III, 1 step w/Platinum Taq 100 SuperScript One Step RT-PCR with Platinum Taq Superscript™ II Reverse Transcriptase Superscript™ II Reverse Transcriptase Superscript™ Reverse Transcriptase SURE® 2 Supercompetent Cells SURE® Competent Cells Swa I Sybr Green PCR Master Mix Sybr Green PCR Master Mix SYBR Green qPCR Syringe fltr-Durapore PVDF/PVC .22µ Syringe fltr-Durapore PVDF/PVC .45µ Syringe fltr-mixed esters of cellulose .22µ Syringe fltr-mixed esters of cellulose .45µ per hour 12/pk 12/pk 12/pk 12/pk 100 µg 1000u 400u 3000u 2000u 1 kg 1 kg 3 mg/ml 10 rxn 50 rxn 10000u 100 rxn 25 rxn 10000u 4 x 10000u 10000u ea 5x0.2ml 200u pk 10 pk 100 rxn 50/pk 50/pk 50/pk 50/pk -TOE OE OE OE OE OE OE OE OE OE OE OE OE OE OE OE OE OE OE OE OE KT T3 RNA Polymerase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase T4 DNA Ligase HC T4 DNA Polymerase T4 DNA Polymerase T4 DNA Polymerase T4 DNA Polymerase T4 DNA Polymerase T4 DNA Rapid Ligation Kit (5 minutes) T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase T4 Polynucleotide Kinase FPLCpure T7 RNA Polymerase TA Cloning Kit® with Dual Promoters (INVaF') 1000u 100u 100u 500u 20000u 100000u 100u 20000u 100u 50u 250u 150u 100u 40 rxn 200u 200u 500u 2500u 100u 200u 1000u 20 rxn University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 130 CH MB MB PC PC PC PC PC PC PC PC PC PC PC PC PC PC PC PC PC PC CH CH TC TC TC CH CH OE OE OE CH CH MB TC TC PC PC PC OE PC PC IM KT KT KT KT KT KT KT KT KT KT KT SV TAE 10X Talon Metal Affinity Resin Talon Superflow Metal Affinity Resin Taq DNA Polymerase Taq DNA Polymerase Taq DNA Polymerase Taq DNA Polymerase Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer Taq DNA Polymerase w/ buffer A Taq DNA Polymerase w/ buffer B Taq DNA Polymerase w/10X Buffer Taq PCR Core Kit TaqMan Universal Master Mix TaqMan Universal Master Mix Taqstart Antibody Taurine TBE 10X TE 10X, pH 7.4 TE, 1X Solution, pH 7.4 TE, 1X Solution, pH 8.0 TEMED TEMED Terminal Deoxy Transferase (TdT) Terminal Deoxy Transferase (TdT) Terminal Deoxy Transferase (TdT) Terrific Broth (Modified)-EZMix® Tetracycline 12.5mg/ml Tfx-10 Transfection Reagent TGF-alpha - Human Recombinant TGF-beta Human Natural Tgo DNA Polymerase Tgo DNA Polymerase Tgo DNA Polymerase Thrombin Protease TITAN™ One-Tube RT-PCR System TITAN™ One-Tube RT-PCR System TNF-a TNT®-SP6 Coupled Transcription/Translation TNT®-SP6/T7 Coupled Transcription/Translation TNT®-T3 Coupled Transcription/Translation TNT®-T7 Coupled Transcription/Translation TNT®-T7 QUICK Coupled Transcription/Translation TNT®-T7/T3 Coupled Transcription/Translation TOPO TA Cloning® Kit (TOP10 Cells) TOPO TA Cloning® Kit (TOP10F' Cells) TOPO TA Cloning® Kit (with LTI 4622 - DH5a Cells) TOPO TA Cloning® Kit Dual Promoter (TOP10F' Cells) TOPO TA Cloning® Kit for Sequencing (Top 10 Cells) Transition to protein free medium 4L 10 ml 25 ml 250u 10x250u 250u 2500u 100u 250u 5000u 100u 500u 100u 500u 100u 100u 250u 100 rxn 200 rxn 2000 rxn 200 tst 1 kg 1L 1L 1L 1L 50 g 5 ml 2500u 500u 300u 6 x 500ml 10 ml 1 ea 50 µg 1 µg 50 rxns 100 rxns 250 rxns 500u 100 rxn 25 rxn 10µg 1 kit 1 kit 1 kit 1 kit 1 kit 1 kit 20 rxn 20 rxns 20 rxns 1 ea 20 rxn per line University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 131 CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH CH MB OE CH TC TC TC TC TC CH CH NR NR CH Tri Reagent Tri Reagent Tris Tris Tris TRIS pH 7.5 TRIS pH 8.0 Tris Base TRIS EDTA Solution 100X Tris Hydrochloride TRIS/Glycine/SDS 10X Buffer Tris-Base Tris-Glycine (10X) Triton® X-100 TRIZMA® Base Molecular Biology Reagent Grade TRIZMA® Base Reagent Grade TRIZMA® Hydrochloride Reagent Grade Trizma-Hcl, 1 M Stock Solution, pH 7.6 Trizma-Hcl, 1 M Stock Solution, pH 8.0 Trizol TRIzol Reagent tRNA Trypan Blue Solution Trypsin - 2.5% (10X) Trypsin-EDTA - 0.05% T - .53mM EDTA Trypsin-EDTA - 0.05% T (1X) Trypsin-EDTA - 0.25% (1X) Trypsin-EDTA - 0.5% (10X) Tryptone Tryptone Peptone TUNEL Enzyme (optimized terminal transferase) TUNEL Label (FITC dUTP + dNTPs) Tween 20 25 ml 100 ml 1 kg 500 g 1 kg 1L 1L 1 kg 1L 500 g 1L 500 g 1L 100 ml 1 kg 1 kg 1 kg 1L 1L 200 ml 100 ml 100 mg 100ML 100 ml 100 ml 100 ml 100 ml 100 ml 500 g 500 g 1 ea 1 ea 100 ml -U- -V- -WCH CH PC PC PC TC TC TC CH IM CO CO CO CO CO CO CO CO CO CO Urea Urea UTP 0.1M Vent® (exo-) Polymerase Vent® DNA Polymerase Versene - 0.02% EDTA in HBSS Vitamin Mixture - 100X vitamin supplement Water Sterile - Tissue Culture Grade Water(Sterile Nuclease Free) Western Blue Stabilized Substrate for AP Wizard™ DNA Clean-up System Wizard™ Lambda System Wizard™ Maxipreps System Wizard™ Megapreps System Wizard™ Midipreps System Wizard™ Minicolumns Wizard™ Miniprep DNA Purification Resin Wizard™ PCR Preps System Wizard™ Plus Miniprep System Wizard™ Plus Miniprep System 500 g 2 kg 25 µmol 200u 200u 1L 100 ml 1L 1L 1 ea 100 col 20 col 10 col 5 col 25 col 250 col 250 ml 50 col 50 col 250 col University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 132 CO CO MS Wizard™ Plus SV Wizard™ Plus SV Wrapping Film (18in x 1000ft) 50 rxn 250 rxn ea -X- -Y- -ZRE RE RE RE RE RE RE CH CH CH RE RE RE RE RE CE CE CE CE CE RE RE FM FM FM FM FM CH CH MB MB MB CH MB KT FM Xba I Xba I Xba I Xba I Xba I Xba I Xba I X-gal X-gal X-gal Xho I Xho I Xho I Xho I Xho I HC XL 10-Gold Ultracompetent Cells XL-1 Blue Electroporation Competent Cells XL10-Gold Ultracompetent Cells XL-1-Blue Supercompetent Cells XL-2 Blue Ultra Competent Cells Xma I Xmn I X-Omat™ AR-2 interleaved film 35x43cm X-Omat™ AR-2 interleaved film 8x10in X-Omat™ AR-5 non-interleaved film 8x10in X-Omat™ Blue 5x7 X-Omat™ Blue 8x10in Yeast Extract Yeast Extract Yeast tRNA Yeastmaker Carrier DNA Yeastmaker Yeast Transformation System YT Broth 2X Zeocin Zero Blunt® TOPO® PCR Cloning Kit (TOP 10 Cells) Zeta-Probe® GT Membrane15x15cm 1500u 1000u 2000u 10000u 3000u 15000u 2000u 1g 250 mg 1g 5000u 25000u 3000u 5000u 2500u 10 x 100µl 5x80µl 5x100ul 5x0.2ml ea 250u 1000u 50 sht 50 sht 50 sht 100/bx 100/bx 500 g 500 g 25 mg ea kit 500 g 1g 20 rxn 15 sht University of Pennsylvania Department of Genetics Core Facilities Catalog 2004 – page 133