Gene Transfer, Targeting
and Therapeutics Core

Scheduling & Rates

Salk Institute for Biological Studies - Gene Transfer, Targeting
and Therapeutics Core - Scheduling & Rates

Scheduling & Rates


Forms

Order from available pre-made GT3 Stocks:

  • Click here to download a request form for aliquots of viral vector reporter stocks and stock cell lines

Order a custom vector production using your transfer construct or one of the transfer constructs available at the core.

  • Click here to see the current list of AAV, FIV, LV, Retro or EIAV transfer plasmids available at no charge at the core (Internal link for Salk researchers only)
  • Click here to download a request form for custom recombinant viral vector production

Recharge Rates


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The Salk GT3 core provides concentrated high titer stocks of pseudotyped lentivirus (HIV-1 and EIAV) and retrovirus (MMLV), E1-deleted Adenovirus (Ad5) and rAAV2 cross-packaged with a range of different capsids.

Rates are given for standard preparations from 12 x 15 cm dishes, which typically yield ~200 ul 108-1010 GC/ml lentivirus or ~3.5 ml of 1011-1013 GC/ml rAAV. Purity is suitable for both in vitro and in vivo applications.



Lentiviral and Retroviral Vectors

Large-scale production and purification of pseudotyped lentiviral and retroviral vectors is performed by transient transfection of 12 x 15 cm dishes followed by ultracentrifugation on a sucrose cushion. Upon request, the sucrose cushion can be replaced with purification using large volume of PSB, which was found to lower the viscosity of the preps.

Medium containing replication-deficient lentiviral or retroviral particles is harvested twice and pooled before filtration and concentration. Concentrated virus is resuspended in HBSS, aliquoted and stored at -80 degrees C.

Custom preps are titrated by serial dilution and transduction of 293T cells. Titer is obtained by qPCR (no reporter) to give infectious titer in integrated genome copies (IU) per mL or, upon request, by FACS (if fluorescent reporter present) to give titer in transforming units (TU) per mL.

C3/SCRM Member External
Consultation $50 per hr $50 per hr
Standard Prep with titration $1200 $2000
Stock reporter virus (10 uL) $80 $140

Click to visit the Salk Intranet for internal pricing>>



rAAV2 vectors

Standard scale production and purification of cross-packaged rAAV vector is performed by transient transfection of 12 x 15 cm dishes followed by purification on a discontinous OptiprepTM gradient. Further polishing by size exclusion is available by request.

Large scale production and purification is performed by transient transfection of 60 x 15 cm dishes followed by purification on two consecutive CsCl gradients. Collected fraction are analysis by separation on SDS gel and silver staining, RI determination and QPCR analysis. The fractions contained highest percentage of full capsids and lowest amount of contaminating proteins are pulled and dialyzed against PBS and PBS/Sorbitol.

Gradient fractions containing purified rAAV infectious particles are aliquoted and stored at -80 degrees C.

Custom rAAV preps are titrated using qPCR to give titer in genome copies (GC) per ml.

C3/SCRM Member External
Consultation $50 per hr $50 per hr
Standard Prep with titration (Iodoxanol) $1200 $2000
Iodoxanol Stock reporter vector (50ul) $80 $130
Large Prep with titration (CsCl) $1700 $2900
CsCl Stock reporter vector (50ul) $160 $270

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G-deleted rabies


Large scale production and purification of G-deleted rabies vectors is performed on BHK21 cells and the vector is amplified folowing initial infection with seed virus.

Purified G-deleted rabies infectious particles are aliquoted and stored at -80 degrees C.

Custom rabies vector preps are titrated using infection of reporter cells and qPCR to give titer in infectious units(IU) per ml.

C3/SCRM Member External
Consultation $50 per hr $50 per hr
Standard Prep from seed with titration $2400 $4000
Stock G-deleted rabies (10ul) $130 $200
Custom Rabies Recovery from DNA $3500 $5850

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Ad5 vectors

Large scale production and purification of E1a/b and E3 deleted Ad5 vectors is by transient transfection of Pac 1 digested Ad5 vector DNA, amplification of vector on 293 cells followed by purification on a discontinous OptiprepTM gradient and polishing using a size exclusion column.

Gradient fractions containing purified Ad5 infectious particles are aliquoted and stored at -80 degrees C.

Custom Ad5 preps are titrated using qPCR to give titer in genome copies (GC) per ml.

C3/SCRM Member External
Consultation $50 per hr $50 per hr
Standard Prep with titration (from stock) $1700 $2800
Standard Prep with titration (from DNA) $2300 $3900
Ad5 stocks $100 $160

Click to visit the Salk Intranet for internal pricing>>