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T98G

Multiporator Transfection Protocol Protocol No. 4308 915.041 11/2000 Cell line T98G, human glioblastoma Transfection with Plasmid pEGFP-N1 (in bidistilled H2O) Electroporation buffer Eppendorf Hypoosmolar Electroporation Buffer (PH) Culture medium IMDM / 10% FCS Cuvette Eppendorf, 2 mm gap width, 400 l Temperature RT (20-25 C) Reference Dr. Ariane Sling Martin-Luther-Universitt Halle Molekulare Neuroonkologie
Magedburger Str. 16 D-06097 Halle/Saale
Phone +49 345 5574050 Fax +49 345 5571412 e-mail: ariane.soeling@medizin.uni-halle.de
  1. Harvest the cells in the exponential growth phase and centrifuge them (for 5-10 minutes, 200 x g, at room temperature).
  2. Resuspend the cells in IMDM / 0.5% FCS, determine the number of cells and centrifuge them (for 5-10 minutes, 200 x g, at room temperature). 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 10'"/>


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