VIRAL VECTOR PLATFORM SERVICE
LENTIVIRUS APPLICATION FORM (REG-VVB-PVV-PNT-001-01.01)
BILLING
REQUESTER:
PI:
INSTITUTION:
ADDRESS:
PHONE NUMBER:
PI's e-MAIL:
DELIVERY (do not fill in if the same as above)
NAME:
INSTITUTION:
SHIPPING ADDRESS:
PHONE NUMBER:
e-MAIL:
Please fill the data below and send the document by e-mail to Naiara Tejados(1)
NUMBER OF BATCHES (2)
Mini-Lenti: Supernatant from 1 P100 packaging-cell transfection
Midi-Lenti: Supernatant from 1 P150 packaging-cell transfection
Maxi-Lenti: Supernatant from 3 P150 packaging-cell transfection
Note: If you need more than one viral vector separate the data of each one with //
TYPE OF VIRAL VECTOR
Shuttle plasmid (name, designation)
Length (in Kbp) of the packable construct
Transgene (type, special features if any)
Reporter gene
Promoter
Generation
2nd
3rd
Self-inactivating
Yes
No
Pseudotype (indicate whenever VSVg is not
the env gene)
VIVEbioTECH - Parque Científico y Tecnológico de Gipuzkoa // P. Mikeletegi 81 // 20009 - San Sebastián (Gipuzkoa) País Vasco (Spain)
www.vivebiotech.com // info@vivebiotech.com // Telf. 943308568 // Fax. 943308222
DNA PURITY OF SHUTTLE PLASMID (PROVIDE BY USER) (3)
260/280
Volume
Concentration
TITRATION (4) (5)
Titration by default
Transduction units (T.U./mL)
Viral particles containing genome (v.g./mL)
Titration on request (at an additional cost)
p24 ELISA
Viral inserted copies (inserts/cell)
CONCENTRATION
Concentration using PEG (available for Mini-Lenti and Midi-Lenti)
Ultracentrifugation (available for Maxi-Lenti)
VIVEbioTECH - Parque Científico y Tecnológico de Gipuzkoa // P. Mikeletegi 81 // 20009 - San Sebastián (Gipuzkoa) País Vasco (Spain)
www.vivebiotech.com // info@vivebiotech.com // Telf. 943308568 // Fax. 943308222
ADDITIONAL NOTES:
1. For any additional information or comments please contact Naiara Tejados (send) who will
pass on any doubts and additional information requests to the production personnel.
2. SCALE OF PRODUCTION
Producer cells: HEK293T. Supernatant per P150 dish: 20 mL.
Mini-Lenti: 1 P100 supernatant. Batches cannot be concentrated by ultracentrifugation.
Clarified supernatant will be delivered. Vectors can be concentrated using PEG on request.
Midi-Lenti: 1 P150 supernatant. Batches cannot be concentrated by ultracentrifugation.
Clarified supernatant will be delivered. Vectors can be concentrated using PEG on request.
Maxi-Lenti: 3 P150 supernatant. Vectors can be concentrated by ultracentrifugation on
request.
3. QUALITY of DNA
Minimum purity values 260/280=1.8. RNA free. A photo of an EtBr or similarly stained
agarose gel will be required (attach as pdf or jpg format). Competent Stbl3 or similar, and
Endofree DNA purification columns are strongly recommended.
DNA must be resuspended in H2O at 1 mg/mL. 30 micrograms of DNA are needed to
transfect one P150 dish.
4. TITRATION methods.
ALL the batches produced at VIVEbioTECH will be titrated and data delivered. Depending on
the specificities of the vectors, titrations will be done by default as follows:
 Whenever the transfer plasmid does encode for a fluorescent reporter that can be
properly excited by the 488 or the 635 nm laser, ALL the batches will be titrated by
transducing indicator HEK293T cells and FACS analysis.
Unit: transduction units per mL (T.U./mL).
These batches can also be titrated by qRT-PCR under request.
 Whenever the transfer plasmid does not encode for a fluorescent reporter, ALL the
batches will be titrated by qRT-PCR.
Unit: viral genomes per mL (vg/mL).
Normally, following our procedures, 1 T.U. is equal to 300-500 viral-genome
containing particles.
 For viruses bearing an antibiotic-resistant gene, inquire for pricing.
 Additionally, titration by integration events by qPCR on genomic DNA or by p24 ELISA
can be provided upon request with additional cost.
Final titers are regularly >1x106T.U./mL for non-concentrated batches and >1x108 T.U./mL
for concentrated stocks.
5. YIELDS.
Final yields depend on the size and specificities of the shuttle plasmid and the transgene.
IMPORTANT INFORMATION
1- All vectors delivered by the platform are for research use only.
2- The generated lentivectors need to be handled and used under BSL-II.
VIVEbioTECH - Parque Científico y Tecnológico de Gipuzkoa // P. Mikeletegi 81 // 20009 - San Sebastián (Gipuzkoa) País Vasco (Spain)
www.vivebiotech.com // info@vivebiotech.com // Telf. 943308568 // Fax. 943308222
GENERAL CONDITIONS
1. Lentiviral stocks will be dispatched within one month following receipt of the transfer plasmid,
under the conditions specified above. The customer will be notified of any delay due to
justified but unforeseeable shortly after realisation.
2. The customer’s material will be safeguarded for a six-month period after the delivery of the
viral stocks. Thereafter it will be destroyed. The customer will be informed of any incident
regarding the quality, damage or loss of the material during storage at VIVEbioTECH, and
this will be duly registered according to VIVEbioTECH Quality Guidelines.
3. VIVEbioTECH ensures the complete confidentiality for the whole process, for both the
reagents and the materials from the customer’s side, and the produced viruses.
4. Production of lentiviral vectors at VIVEbioTECH is based on the following reagents and
procedures: psPAX2 (Addgene plasmid 12260) and pMD2.G (Addgene plasmid 12259) will
be used for second-generation lentivirus production, and pMD2.G (Addgene plasmid 12259),
pMDLg/pRRE (Addgene plasmid 12251) and pRSV-Rev (Addgene plasmid 12253) will be
employed for third-generation lentivirus production. All these materials were kindly
transferred to Addgene by Dr. Didier Trono. We strongly encourage customers to refer to this
source of the helper plasmids and the reference “A third generation lentivirus vector with a
conditional packaging system” J. Virol. 1998 Nov. 72 (11): 8463-71 in the Materials and
Methods section whenever data regarding the lentivirus is published or shared.
5. VIVEbioTECH would appreciate the inclusion of “VIVEBIOTECH SL” as the manufacturer of
the lentivirus in any related publications or research projects.
VIVEbioTECH - Parque Científico y Tecnológico de Gipuzkoa // P. Mikeletegi 81 // 20009 - San Sebastián (Gipuzkoa) País Vasco (Spain)
www.vivebiotech.com // info@vivebiotech.com // Telf. 943308568 // Fax. 943308222