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COS-7

Multiporator Transfection Protocol Protocol No. 4308 915.022 11/1999 Cell line COS-7, monkey, kidney Transfection with Plasmid RSV/lacZ (in bidistilled H2O) Electroporation buffer Eppendorf Hypoosmolar Electroporation Buffer (PH) Culture medium DMEM / 10% FCS Cuvette Eppendorf, 2 mm gap width, 400 l Temperature 4 C Reference Dr. Helena Edlund Department of Microbiology University of Umea Umea Sweden
Phone +46 90 7856701 Fax +46 90 772630 e-mail: Helena.Edlund@micro.umu.se
  1. Harvest the cells in the exponential growth phase and centrifuge them (for 5-10 minutes, 200 x g, at 4 C).
  2. Resuspend the cells in DMEM / 0.5% FCS, determine the number of cells and centrifuge them (for 5-10 minutes, 200 x g, at 4 C). Remove supernatant.

    Note: The overall incubation time in the Eppendorf Electroporation Buffer must not exceed 30 minutes to guarantee a successful electroporation!

  3. Resuspend the cells in Hypoosmolar Electroporation Buffer. When doing so, set the cell concentration to 1 x 106 cells/ml.
  4. Add and mix plasmid DNA (5 g/ml final concentration, in bidistilled
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