Difference between revisions of "Team:Paris Bettencourt/Protocols/Heat Shock transformation"

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     <td><b>Heat Shock transformation protocol</b><br/>
 
     <td><b>Heat Shock transformation protocol</b><br/>

Latest revision as of 17:34, 14 August 2015

Heat Shock transformation protocol
  • Thaw frozen chemically competent cells (20µL aliquots) on ice for 10min.
  • add 2µL of ligation product (or 0.5µL of miniprep) and incubate the cells 30sec at 42°C.
  • put the cells back on ice for 2min.
  • add 200µL of LB to the cells and incubate 2 hours at 37°C.
  • plate the cells on LB + erythromycin (150µg/mL) or LB + erythromycin (10µg/mL), incubation at 37°C overnight.