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- For one 24-well:
- Combine 36μL serum-free media and 4μL Lipofectin in Eppendorf.
- Incubate for 30 minutes at room temperature.
- Combine 30μL serum-free media and 10μL 20μM sense or antisense oligo in another Eppendorf.
- Incubate for 5 - 10 minutes at room temperature.
- Add Lipofectin dilution to sense/antisense dilution.
- Incubate for 15 minutes at room temperature.
- Add 120μL serum-free media.
- Remove media from well and add 200μL of the above mixture to each well.
- After 4 hours, add serum to give a final concentration of 10% per well.
- Culture for the required amount of time.
- For a 96-well, divide the above volumes by 5, for a 6-well, multiply the above volumes by 5.