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TransPass D1 Protocol 1: Transfection in the presence of serum |  | | TransPass D1 Protocol 1: Transfection in the presence of serum |
Introduction
The amounts below are given for a 12-well plate format. Use Table 1 to adjust the reagent volumes for other plate sizes.
Protocol
- Cultures should be grown in 0.5 ml medium containing 5% serum. Cells to be transfected should be 70-80% confluent at the time of transfection.
- Mix 0.5-1.2 µg plasmid DNA in 100 µl serum-free medium.
- Briefly vortex the tube of TransPass D1 Transfection Reagent and pipet 0.5-3.2 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.
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