Transformation

From 2006.igem.org

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Revision as of 16:25, 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
• leave on ice for 10min
• add 2uL DNA to competent cell (label)
• mix with pipette
• sit 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
• leave in incubator 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 plates to dry
• leave in incubator overnig