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oncentration to 2 x 106 cell/ml.
  • Add and mix plasmid DNA (15 g/ml final concentration, in bidistilled H2O).
  • Transfer 400 l cell suspension into electroporation cuvettes (2 mm gap width). The cell suspension must be free of air bubbles.
  • Electroporation:

    Mode Eukaryotes Voltage (V) 180 V Time constant (T) 100 s No. of pulses (n) 1
  • After the pulse, allow the cell suspension to stand in the cuvette for 10 minutes at room temperature.
  • Carefully transfer the cell suspension from the cuvette into 3 ml M3 / 5% FCS, and cultivate them in a 35 mm culture dish of a 6-well plate.

    Detection methods for transfection:

    The expression of EGFP driven by the insect actin promotor can be detected clearly after 24-48 hours by FACS analysis
    or under a fluorescence microscope. Result: Survival rate: 70-80% Transfection rate (stable): 6-9% based on the number of surviving cells. Results were measured 24-48 hours after transfection.


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