(Jennifer Kennell, 2/05)
Materials needed: - BOSC packaging cells (Rack 5, box A in room 7615)
- pVSVG vector (R77)
- retroviral vector of choice (pLXSN, etc)
- 8 mg/mL Polybrene
- 0.45 uM syringe filters
- syringes
- Transiently transfect BOSC packaging cells with 5 µg of pVSVG and 5 µg of retroviral vector using calcium-phosphate precipitation. Shock (glycerol/PBS) and feed cells 4-6 hours after transfection.
- Feed transfected cells 24 hours later
- 48 hours after transfecting, collect media from BOSC cells and filter through 0.45 µM syringe filter. Add polybrene to final concentration of 8 µg/mL
- Transfer media containing virus to cells you want to infect
- Split cells into selection media once cells get to correct density.
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