Transformation

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* incubate overnight
* incubate overnight
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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

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