Team:Evry/Notebook/Protocols/Transformation
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Revision as of 19:52, 17 October 2014
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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