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oosmolar Electroporation Buffer. When doing so, set the cell concentration to 2 x 107 cells/ml.
  • Add and mix plasmid DNA (25 g/ml final concentration, in bidistilled H2O).
  • Transfer 800 l cell suspension into electroporation cuvettes (4 mm gap width). The cell suspension must be free of air bubbles.
  • Electroporation:

    Mode Eukaryotes Voltage (V) 400 V Time constant (T) 50 s No. of pulses (n) 2
  • After the pulse, allow the cell suspension to stand in the cuvette for 5 minutes at room temperature.
  • Carefully transfer the cell suspension with a pasteur pipette from the cuvette into 5 ml RPMI / 10% FCS, and cultivate them in a little culture flask.
    • Detection methods for transfection:
    Selection and limiting dilution assay for stable transfection after 48 hours. Result: Survival rate: 85% Transfection rate: 40% based on the number of surviving cells. Results were measured 48 hours after transfection.


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