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Team:Newcastle/E. coli Competence
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Revision as of 13:43, 29 July 2010 by Shethharsh08 (Talk | contribs)
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Competent cells preparation
Materials required
- 300ml of LB broth
- 1/20 volume of an overnight culture
Procedures
- Inoculate 300 ml of LB broth with 1/20 volume of an overnight culture of the desired strain.
- Grow the cell at 37°C (with shaking platform so as to mix the media equally amongst the cells)
- Next Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes
- Resuspend the pellet in 100 ml of ice cold TFBII
- Aliquot 200 µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice
- Store at -80°C
   
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