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Team:TU Darmstadt/Protocols/Electroporation
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== Electroporation == | == Electroporation == | ||
| + | === About === | ||
| + | Electroporation is a [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Bacterial_transformation 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 ddH<sub>2</sub>O to reduce the conductivity | ||
Revision as of 16:27, 16 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
