1. Seed cells overnight before the day of transfection.

2. For each 35 mm plate or each well of a 6-well plate, add 100 ml serum-free medium to a sterile tube. Add 3-6 ml of FuGENE 6 reagent directly into the media. Always add the serum-free medium first. Tap to mix.

3. Add 1-2 mg DNA to the mix. Tap gently to mix.

4. Incubate for 15 minutes at room temperature.

5. Add dropwise the DNA mixture to the culture cells. Distribute the mixture around the dish by gently swirling.

6. Return the cells to the incubator.

7. Replace medium the next day with fresh medium.