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Team:Newcastle/E. coli Competence
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RachelBoyd (Talk | contribs) (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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{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
| - | + | =Competent cells preparation= | |
| - | #Inoculate | + | ==Procedures== |
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| + | #Inoculate 300 ml with 1/20 volume of an overnight culture of the desired strain. | ||
#Grow the cell at 37°C (with shaking) | #Grow the cell at 37°C (with shaking) | ||
#Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes | #Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes | ||
#Resuspend the pellet in 100ml of ice cold TFBII | #Resuspend the pellet in 100ml of ice cold TFBII | ||
| - | #Aliquot | + | #Aliquot 200 µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice |
#Store at -80°C | #Store at -80°C | ||
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| + | {{Team:Newcastle/footer}} | ||
Revision as of 14:55, 28 July 2010
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Competent cells preparation
Procedures
- Inoculate 300 ml 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 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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