Dharmafect transfection (siRNAs)

Tuesday: 19.06.18.

siRNA preparation

All steps at RT, mix by pipetting carefully up and down.

  1. Tube 1: mix siRNAs (200ng/µl), add 2.5 µl (500ng, for a 35mm well) to 197.5 µl of gentamycin (GA) free Lonza
  2. Tube 2: add 2 µl of Dharmafect to 198 µl GA free Lonza
  3. incubate 5’
  4. mix tubes 1 & 2
  5. incubate 20’

Transfection

  1. Use cells at ≈50% confluency
  2. Change media to Lonza without GA, 1 ml per well
  3. Add siRNA mix to cells (400 µl / well), incubate 4h
  4. Change media to normal Lonza
  5. Assay cells next day (can be trypsinized, plated etc)