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TransPass COS/293: Transfection Protocol


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


  1. Cultures should be grown in 0.5 ml medium containing 10% serum. Cells to be transfected should be 70-080% confluent at the time of transfection.
  2. Mix 1.5 µg plasmid DNA in 100 µl serum-free DMEM medium.
  3. Gently mix TransPass COS/293 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.
  4. Incubate at room temperature for 20-30 minutes to form the transfection complex.
  5. Add the transfection complex mixture to cells. Rock the plate gently in order to evenly disperse the complex mixture.
  6. Return the plate to the incubator and incubate 24-72 hours before assaying.
  7. Replace medium as needed to maintain healthy cells.




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