Lipofection

Dominik Haudenschild, 10-28-94 at CBRC

1) Precipitate 10ug DNA

2) Move to TC Hood. Decant Supernatant, rinse DNA with 80% then 100%

ethanol using sterile procedure. Allow to dry.

3) Resuspend DNA in 50ul Sterile water by repeated pipetting.

4) Put DNA into 15ml polystyrene tube. (clear plastic)

5) Add 50ul Lipofectin. Wait 15 minutes, solution will get cloudy.

6) Wash 50%-80% confluent cells twice with 3ml Opti-MEM.

7) Give cells 5ml Opti-MEM

8) Add Lipofectin/DNA dropwise.

9) Incubate overnight in TC incubator.

10) Replace Opti-MEM with 10ml normal cell growth medium.

11) Allow cells to grow until very confluent (3-4 days) then harvest

medium.