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TransPass D2 Protocol 1: Transfection in the presence of serum |  | | TransPass D2 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 5-10% serum. Cells to be transfected should be 70-80% confluent 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.

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