Difference between revisions of "Team:Paris Bettencourt/Protocols/Electroporation"

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     <td><b>Electroporation Protocol</b><br>
 
     <td><b>Electroporation Protocol</b><br>

Latest revision as of 17:34, 14 August 2015

Electroporation Protocol
  • Thaw electrocompetent cells on ice
  • Add 2µL of ligation product or 0.5µL of native plasmid to the cells
  • Transfer the cells in an 0.2mm electroporation cuvette
  • put the cuvette in the electroporation device and pulse the cells at 2.5kV, 200 Ohms and 25µF
  • add 200µL of LB right after pulsing
  • recover 2 hours at 37°C
  • plate 200µL and 50µL of the cells on LB + erythromycin (200µg/mL), incubate overnight at 37°C