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Team:TU Darmstadt/Protocols/Electroporation
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Latest revision as of 21:14, 26 September 2012
Contents |
Electroporation
About
Electroporation is a transformation method. Genetic material is transfered into competent cells by an electric pulse.
Procedure
- Defrost a stock of competent cells on ice
- Mix it with 1 µL of plasmid
- 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
- Pulse
- Add 1 mL of DYT medium to cell suspension directly after the pulse and incubate at 37°C for 1 h
- 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
