Parque Científico y Tecnológico de Gipuzkoa. P Mikeletegi 81
20009 San Sebastián
Guipuzcoa, Pais Vasco España
Telf. 943309064
Fax. 943308222
VIRAL VECTOR PLATFORM SERVICE
APPLICATION FORM LENTIVIRUS
REF:
CUSTOMER:
PI:
INSTITUTION:
ADDRESS:
SHIPPING ADDRESS:
PHONE NUMBER:
e-MAIL:
BILLING ADDRESS:
Please fill the data below and send the doc by e-mail to Naiara Tejados(1)
- Number of batches (2)
 Mini-Lenti: Supernatant from 1 P100 packaging transfected cells
 Mind-Lenti: Supernatant from 1 P150 packaging transfected cells
 Maxi-Lenti: Supernatant from 3 P150 packaging transfected cells
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- 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 (3)
2nd 3rd 
 Self-inactivating
yes
no 
 Pseudotype (indicate whenever VSVg is not the env gene)
- DNA purity of shuttle plasmid (provide by user) (3)
 260/280
 Volume
 Concentration
- Titration (4) (5)
Titration by default
 Transduction units (T.U./mL)
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 Viral particles containing genome (v.g./mL)
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Titration under request (with additional cost)
 p24 ELISA
 Viral inserted copies (inserts/cell)
- Concentration
 Concentration using PEG (available for Mini-Retro and Midi-Retro)
 Ultracentrifugation (available for Maxi-Retro)
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Parque Científico y Tecnológico de Gipuzkoa. P Mikeletegi 81
20009 San Sebastián
Guipuzcoa, Pais Vasco España
Telf. 943309064
Fax. 943308222
Additional notes
1. For any additional information or comments please contact Naiara Tejados (send) who
will transfer 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: 1P100 SN. Batches cannot be concentrated by ultracentrifugation. Clarified
supernatant will be delivered. Vectors can be concentrated using PEG under request.
Midi-Retro: 1 P150 SN. Batches cannot be concentrated by ultracentrifugation. Clarified
supernatant will be delivered. Vectors can be concentrated using PEG under request.
MaxiLenti: 3P150 SN. Vector can be concentrated by ultracentrifugation under 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 vector titration will be done by default as follows:
 Whenever the transfer plasmid do encode for a fluorescent marker, ALL the
batches will be titrated by transduction of indicator HEK293T cell and FACS
analysis (included in the price). Fluorescent reporters´ absorption: 488–635 nm;
emission: 530-670 nm. For viruses bearing an antibiotic resistance gene inquire
for pricing.
Unit: transduction unit per mL (TU/mL)
Under request these batches can be also titrated by qRT-PCR (see below) with
an additional cost (see price)
 Whenever the transfer plasmid does not encode for a fluorescent reporter, ALL
the batches will be titrated by qRT-PCR on purified vector particles (included in
the price).
Unit: viral genomes per mL (vg/mL).
Regularly following our procedures 1T.U. is equal to 300-500 viral genome
containing particles.
Additionally, titration by integration events by qPCR on genomic DNA or by p24 ELISA
can be provided upon request with an additional cost (see price).
5. YIELDS.
Final yield depends on the size and specificities of the shuttle plasmid and transgen as
well.
Final titers are regularly >1x106T.U./mL for non-concentrated batches and >1x108
T.U./mL for concentrated stocks.
1- IMPORTANT INFORMATION
2- All vectors delivered by the platform are for research use only.
3- The generated lentivectors need to be manipulated and used in BSL-II.
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Parque Científico y Tecnológico de Gipuzkoa. P Mikeletegi 81
20009 San Sebastián
Guipuzcoa, Pais Vasco España
Telf. 943309064
Fax. 943308222
GENERAL CONDITIONS
1- Lentiviral stocks will be dispatched before one month upon receipt of the transfer plasmid
under the conditions specified above. Any delay due to due to justified but unforeseeable
cause will be notified to the customer shortly after realization.
2- The customers´ material will be safeguarded for a six months period after the delivery of
the viral stocks. Thereafter it will be destroyed. Any incident regarding the quality,
damage or loss of the material along the storage at VIVEbioTECH will be informed to the
customer and properly registered according to the Quality Guidance of VIVEbioTECH.
3- VIVEbioTECH ensures the complete confidentiality along the whole process of both the
reagents and material from the customers´ side and the produced virus as well.
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 were kindly
transferred by Dr. Didier Trono to Addgene. We strongly encourage the customers to
refer 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
Material and Methods section whenever data will be published or shared.
5- VIVEbioTECH also acknowledges the indication of “VIVEBITECH SL“ as the
manufacturer source of the lentiviruses employed in research projects
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