Transformation
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Revision as of 16:29, 28 June 2006
iGEM 2006 Lab Training Session I: Transformation
Key words: [http://en.wikipedia.org/wiki/Transformation_%28genetics%29 transformation], [http://en.wikipedia.org/wiki/Competent_cells competent cells]
Procedure
Preparation: competent cells
For each plasmid:
- take competent cells from -80C freezer
- thaw on ice for 10min
- add 2uL DNA to competent cell (label)
- mix with pipette
- sit on ice for 30min
- get a beaker of exactly 42C water from hot water tap
- submerge tubes in hot water bath for exactly 90s
- sit on ice for 5min
- add 1mL LB
- incubate for 1h
- spread 500uL on Petri plate labelled with appropriate antibiotics
- Dip a glass rod with ethanol and flame it to spread cells in plates
- wait for plate to dry
- incubate overnight
