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doing so, set the cell concentration to 5 x 106 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) 530 V Time constant (T) 50 s No. of pulses (n) 2
  • After the pulse, allow the cell suspension to stand in the cuvette for 5-10 minutes at room temperature.
  • Carefully transfer the cell suspension from the cuvette to 5 ml RPMI 1640 / 0.5% FCS, and centrifuge it for 5-10 minutes, 200 x g, at room temperature. Remove supernatant.
  • Resuspend the cells in 1 ml RPMI 1640 / 10% FCS, and cultivate them in a 35 mm culture dish.
    • Detection methods for transfection:
    The expression of the plasmid pEGFP-N1 can be detected clearly after 24-48 hours with the aid of FACS analysis or under a fluorescence microscope. Result: Survival rate: 143% Transfection rate: 84% based on the number of surviving cells
    120% based on the initial number of cells used for the experiment Results were measured 48 hours after transfection.


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