TransPass HeLa: Transfection Protocol
Overview
The amounts below are given for a 12-well plate format. Use Table 1 to adjust the reagent volumes for other plate sizes.
Protocol
- Plate cells in complete growth medium containing 10% serum and no antibiotics/antimycotics at an appropriate density, so that they will reach 70-80% cell density (plasmid transfection) or 40-60% (siRNA alone or plasmid and siRNA transfection) at the time of transfection.
- Add 4 µl of TransPass HeLa Transfection Reagent to 100 µl of serum-free medium (e.g., DMEM/high glucose) in a sterile tube (one per sample well) and mix well.
- Incubate at room temperature for 15 minutes.
- Add 1-3 µg of plasmid DNA and/or 1.5-30 pM of siRNA to the reagent-medium mixture.
- Incubate at room temperature for 15 minutes.
- Add the transfection complex mixture to the well, and evenly disperse the complex mixture by gently rocking the plate.
- Return the plate to the incubator and incubate cells overnight.
- Replace the transfection medium with complete medium (i.e. 10% FBS-DMEM) the next day and incubate 24-72 hours before assaying.