Team:Newcastle/E. coli Competence
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(New page: {{Team:Newcastle/mainbanner}} ==Competent cells preparation== #Inoculate 300ml with 1/20 volume of an overnight culture of the desired strain. #Grow the cell at 37°C (with shaking) #Ch...)
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(New page: {{Team:Newcastle/mainbanner}} ==Competent cells preparation== #Inoculate 300ml with 1/20 volume of an overnight culture of the desired strain. #Grow the cell at 37°C (with shaking) #Ch...)
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Revision as of 12:43, 28 July 2010
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Competent cells preparation
- Inoculate 300ml with 1/20 volume of an overnight culture of the desired strain.
- Grow the cell at 37°C (with shaking)
- Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes
- Resuspend the pellet in 100ml of ice cold TFBII
- Aliquot 200µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice
- Store at -80°C