The amounts below are given for a 12-well plate format. Use Table 1 to adjust the reagent volumes for other plate sizes.
- Plate cells (in complete growth medium containing 5-10% serum and no antibiotics/antimycotics) at an appropriate density so that they will reach 70-80% confluence at the time of transfection.
- Mix 1.5 µg plasmid DNA in 100 µl serum-free medium.
- Gently mix TransPass D2 Transfection Reagent prior to pipetting. Do not vortex. Add 1.5-4 µl to the DNA/medium mix from step 2. Mix gently by flicking the tube.
- Incubate at room temperature for 20-30 minutes to form the transfection complex.
- Add the transfection complex mixture to cells. Rock the plate gently in order to evenly disperse the complex mixture.
- Return the plate to the incubator and incubate 24-72 hours before assaying.
- Replace medium as needed to maintain healthy cells.