Retroviral Transduction 2

[Luke]

Retroviral Transduction 2

1.   Coat plates with RetroNectin (recombinant Fibronectin) at final 12.5ug/mL in PBS for 1.5 hours
2.   Optional: Block using 2%BSA in PBS for 30 minutes
3.   Add 3mL of overnight culture at 3-4million cells/mL density to each well of 6-well plate
4.   Add protamine sulfate or polybrene to each well at 4ug/mL final (including viral supe)
5.   Add 2mL viral supernatant
6.   Centrifuge at 2000RPMs for 90 minutes at 30 degrees
7.   Incubate overnight at 37 5% CO2
8.   Use directly or change ? medium with new.