Fast Microbial Identification Sample sheet Client Invoice Department Your Purchase Order number #: Name of sender: Administrator: Institute / Company: Invoice department: Department: Invoice address: Shipment address: Zip/City: Zip/City: Country: Country: e-mail address: e-mail address: Phone number: Phone number: VAT #: Fax number: Other contact person: e-mail address: Quotation #: Year contract #: ORDER (please use the sample sheet) THE FOLLOWING OPTIONS ARE AVAILABLE, PLEASE INDICATE YOUR CHOICE ON THE SAMPLESHEET: Identification of unknown bacteria: - BC-304F: MicroSEQ® 500bp. The validated MicroSEQ kits and the validated MicroSEQ databases will be used for the identification. BC-308F: Custom ID <700bp. Our own custom 16S kit will be used and the consensus sequences will be compared to an internal database or to a public database. BC-309F: Custom ID <1400 bp. Our own custom 16S kit will be used and the consensus sequences will be compared to an internal database or to a public database. BC-310F: Maldi-TOF. A protein spectrum created by the Maldi-TOF system will be compared to the validated VITEK MS IVD database. Identification of unknown fungi: - BC-306F: MicroSEQ® D2-LSU. The validated MicroSEQ kits and the validated MicroSEQ databases will be used for the identification. BC-307F: ITS-region. Our own custom ITS kit will be used and the consensus sequences will be compared to a public database. BC-310F: Maldi-TOF. A protein spectrum created by the Maldi-TOF system will be compared to the validated VITEK MS IVD database. (This service cannot be used for moulds) RESULTS Depending on the services requested the results will be send only by email or by email and regular mail. BC-304F, BC-306F and BC-307F will receive the results by email and by regular mail. BC-308F, BC-309F and BC-310F will receive the results only by email. Version: 2013-01 F089-MICRO -023 Page 1 of 4 Fast Microbial Identification Sample sheet I accept the general terms and conditions of BaseClear* Name : ______________________ Signature : ______________________ Date :__ / __/ ____ To be filled out by BaseClear: Project Code : __________________ Date of registration : _ _ / _ _ / _ _ _ _ *Our General Terms and Conditions apply to all projects BaseClear carries out and can be downloaded from our website (www.baseclear.com/labservices/terms_conditions). SAMPLES We can only process live Class I, non-pathogenic micro-organism that are provided to us according to Dutch regulations. In brief, this means that bacteria or fungi should be delivered on petri-dishes or on slants that are sealed with parafilm. Sealed dishes/ slants should be wrapped in absorbing tissue, and enclosed in an unbreakable and watertight cover. Please make sure that you clearly indicate the sample names on the petridishes. If you suspect the micro-organisms to be pathogenic please contact us beforehand so we can discuss alternative means of transportation. TERMS OF DELIVERY Fast Bacterial identification using the validated MicroSEQ® system (BC-304F) DNA-isolation, PCR, sequencing and reporting will be performed using MicroSEQ validated protocols, kits and database. Approximately 500 base pairs of the 16S rRNA gene will be amplified and sequenced on both strands. A full report including the homology search to the MicroSEQ validated database and sequence data will be mailed to the client within 1 working days (when samples arrive before 15.00h). The reported result is the most likely sample identification. However, the client has to make the final conclusion to the identity of the sample. Fast Bacterial identification using custom protocols (BC-308F and BC-309F) We offer two possibilities for analysis of bacterial samples using our custom kits. For a custom ID <700bp (BC-308F) approximately 600 base pairs of the 16S rRNA gene will be amplified and sequenced on both strands. A full report including the homology search in an online database and sequence data will be mailed to the client within 2 working days. For a custom ID <1400bp (BC-309F) approximately 1400 base pairs of the 16S rRNA gene will be amplified and sequenced on both strands. A full report including the homology search in an online database and sequence data will be mailed to the client within 2 working days. Because of sequence length a complete search (~1.4 kb) is more reliable than a partial search. The reported result is the most likely sample identification. However, the client has to make the final conclusion to the identity of the sample. Fast Fungal identifiation using the validated MicroSEQ® system (BC-306F) DNA-isolation, PCR, sequencing and reporting will be performed using MicroSEQ validated protocols, kits and database. For fungal samples, the D2-LSU gene (BC-306F) is amplified and sequenced on both strands using validated MicroSEQ® kits and protocols. The length of the consensus sequence is approximately 200-300 bp long. A full report including the homology search to the validated MicroSEQ database and sequence data will be mailed to the client within 1 working days(when samples arrive before 15.00h). The reported result is the most likely sample identification. However, the client has to make the final conclusion to the identity of the sample. Fast Fungal identification using custom protocols (BC-307F) Alternatively, samples can be analysed by amplification and sequencing of the internal transcribed spacer (ITS) genes (BC-307F) using custom kits and protocols, and blasting in public databases. The consensus sequence length is about 1400 bp long. A full report including the homology search in an online database and sequence data will be mailed to the client within 2 working days. The reported result is the most likely sample identification. However, the client has to make the final conclusion to the identity of the sample. Version: 2013-01 F089-MICRO -023 Page 2 of 4 Fast Microbial Identification Sample sheet Bacterial or yeast samples using the Maldi-TOF (BC-310F). It is not possible to requests this service for mould samples, only bacterial and yeast samples will be identified using the Maldi-TOF. Maldi-TOF identification is achieved by obtaining spectra using the Maldi-TOF technology (Matrix Assisted Laser Desorption Ionization Time-of-Flight) and analyzing the spectra with the VITEK MS database. The peaks from these spectra are compared to the characteristic pattern for a species, genus, of family of micro-organism, thus resulting in an identification. If the Maldi-TOF results do not meet BaseClear quality standard, the samples will automatically be identified using BC-308 at no extra charge. Strain comparison (BC-360) Additionally, generated sequencing results can be compared to each other in order to detect differences between strains and visualize them (at additional costs). A pdf report, including visual comparison of the DNA sequences of the strains and marked differences will be delivered. SmartGene database (BC-314) BaseClear has also access to the validated web-based SmartGene databases. Identification of a sample is achieved by comparing the consensus sequence against the constantly updated reference databases. The 16S rDNA Centroid Reference Database is a quality- controlled 16S rDNA Reference Database. The Centroid database is built using a proprietary algorithm, to determine the most representative sequence entry for each bacterial species; searching a sample sequence against the Centroid dataset allows to visualize in a convenient manner the differences of the best matching species Centroid and the other closely related species, for easier discrimination. Version: 2013-01 F089-MICRO -023 Page 3 of 4 Fast Microbial Identification Sample sheet Bacterial identification No. Fungal identification Sample name 1. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 2. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 3. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 4. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 5. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 6. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 7. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 8. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 9. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF 10. MicroSEQ® 500bp Custom ID <700bp Custom ID <1400 bp Maldi-TOF Strain comparison (Y/N) Use SmartGene Database*** (Y/N) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) MicroSEQ® D2-LSU ITS-region Maldi-TOF (only for yeast samples not for mould samples) ***For MicroSEQ samples the use of the SmartGene database is free of charge. For custom ID services the use of the SmartGene databases will be at additional cost. Remarks: Version: 2013-01 F089-MICRO -023 Page 4 of 4