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Multiporator Transfection Protocol Protocol No. 4308 915.046 11/2001 Cell line L2, Schneider cells, Drosophila Transfection with pActin-SV-EGFP Electroporation buffer Eppendorf Electroporation Buffer with 200 mOsmol/kg Culture medium M3 medium / 5% FCS (heat inactivated) Cuvette Eppendorf, 2 mm gap width, 400 l Temperature RT (20-25 C) Reference Dr. Maren Mieth Max-Delbrueck-Centrum fr Molekulare Medizin Robert-Rssle-Str. 10 D-13092 Berlin-Buch Phone +49 30 94062553

1. Harvest the cells in the exponential growth phase and centrifuge them (for 5 minutes, 200 x g, at room temperature).
2. Resuspend the cells in M3 / 0.5% FCS, determine the number of cells and centrifuge them (for 5 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 Electroporation Buffer with 200 mOsmol/kg (= 42% Eppendorf Hypoosmolar Electroporation Buffer + 58% Eppendorf Isoosmolar Electroporation Buffer). When doing so, set the cell c
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