From 2014.igem.org
Transformation of Pseudovibrio
- Place electroporation tanks in ice for 10min
- Take samples of plasmids (25ng-50ng/µL) /!\ Keep them in ice /!\
- Take sample of competent cells /!\ Keep them in ice /!\
- Place 1µL of plasmid in the sample of competent cells
- Transfer the full volume obtained in the electroporation tank
- Place in the electroporator and pulse at 2000V
NB: The optimal pulse length is between 5 and 6ms.
- Add 1mL of MB 1X in the 30 seconds following the transformation
- Incubate between 2h and 3h at 30°C with shaking
- Centrifuge to concentrate all cells in the pellet
- Discard the supernatant
- Sowed the pellet on selective plates of MB 1X
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