Team:TU Darmstadt/Protocols/Electroporation

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Contents

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
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