Team:TU Darmstadt/Protocols/Electroporation

Electroporation

About

Electroporation is a transformation method. Genetic material is transfered into competent cells by an electric pulse.

Procedure

1. Defrost a stock of competent cells on ice
2. Mix it with 1 µL of plasmid
3. Transfer the mixture in an electroporation cuvette
   • Use the following settings for electroporation: 200 Ohm resistance, 2.5 kV current , 2 µF capacity and 2 mm capacitor plate distance
4. Pulse
5. Add 1 mL of DYT medium to cell suspension directly after the pulse and incubate at 37°C for 1 h
6. Finally the cells are ready to be crossed out

Notes

• Important: Use only clean cuvettes for electroporation and wash them very carefully
• Make sure that the cuvettes are perfectly dry to reduce the risk of a shortcut
• Impurities in the plasmid may lead to an arc, if this does occur add small volumes of ddH₂O to reduce the conductivity