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StableTransfection

Stable Transfection (Electroporation)

Outline:

Electroporation can be used for both transient and stable transfection of mammalian cells. Cells are placed in suspension in an appropriate electroporation buffer and put into an electroporation cuvette. DNA is added, the cuvette is connected to a power supply, and the cells are subjected to a high-voltage electrical pulse of defined magnitude and length. The cells are then allowed to recover briefly before they are placed in normal growth medium.

Supplies & Equipment:

Reagents:

Procedure:

  1. DNA preparation
  2. Electroporation
  3. Selection (e.g. with G418)
  4. Cloning

Time Required: five weeks