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From 2014.igem.org

Transformation of competent cells by electroporation

1. Cells preparation:

With the precultures made the day before, add 200µl of bacteria in 15ml of LB medium and add the appropriate antibiotic in a Falcon tube.

Shake vigorously at 30°C until the OD600nm reaches 0,6.

Then, place in an ice bath for 10 minutes and centrifuge the cells for 10 min at 4,000 rpm at 4 °C.

Resuspend the cell pellet in the same initial volume of Glycerol 10% cold and sterile. Centrifuge 5 minutes at 4000rpm at 4°C.

Then, centrifuge the cells for 10 min at 4,000 rpm at 4 °C, and resuspend the cell pellet in the same initial volume of Glycerol 10% cold and sterile. Centrifuge 5 minutes at 4000rpm at 4°C

Then, centrifuge the cells for 10 min at 4,000 rpm at 4 °C.

Resuspend the cell pellet in 100µl of Glycerol 10% cold and sterile.