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
Click to visit the Salk Intranet for internal pricing>>
The Salk GT
3 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 10
8-10
10 GC/ml lentivirus or ~0.5 ml of 10
11-10
13 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 |
$1275 |
$2100 |
| Stock reporter
virus (5 uL) |
$50 |
$75 |
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 (Iodixanol) |
$1275 |
$2100 |
| Iodixanol Stock reporter vector (25ul) |
$55 |
$80 |
| Large Prep with titration (CsCl) |
$1800 |
$3050 |
Click to visit the Salk Intranet for internal pricing>>
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.
Due to recent changes in the Export Administration Regulations regarding rabies vectors, we are unable to distribute outside of the United States any reagents (viral vectors, plasmids, or cell lines) that contain any portion of the rabies genome.
|
C3/SCRM Member |
External |
| Consultation |
$50
per hr |
$50
per hr |
| Standard Prep from seed with
titration |
$2550 |
$4200 |
| Stock G-deleted rabies (5ul) |
$70 |
$105 |
| Custom Rabies Recovery from DNA |
$3700 |
$6200 |
Click to visit the Salk Intranet for internal pricing>>
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 Optiprep
TM 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) |
$1800 |
$3000 |
| Standard Prep with
titration (from DNA) |
$2450 |
$4100 |
| Ad5 stocks |
$105 |
$170 |
Click to visit the Salk Intranet for internal pricing>>
