Team:Bielefeld-CeBiTec/Protocols

iGEM Bielefeld 2015


Protocols



Transformation via Electroporation

  • Thaw 50 µl electrocompetent E. coli cells on ice, dilute with icecold 50 µl glycerine (10%) if necessary
  • Add 0.5-5 µl plasmid to 50 µl electrocompetent cells
  • Store cells on ice for 1 minute
  • Electroporate at U = 2.5 kV, C = 25 µF, R = 400 Ω
  • Transfer transformation reaction to 450 µl SOC-Medium and incubate 1 h at 37 °C
  • Plate on selective LB-Medium
  • Incubate over night at 37 °C

Transformation via Heat Shock

  • Thaw 100 µl chemo competent E. coli cells on ice
  • Add 0.5-5 µl plasmid to 100 µl chemocompetent cells
  • Store cells on ice for 10-30 min on ice
  • Heat shock for 90 seconds at 42 °C
  • Store reaction on ice for 60 seconds
  • Optional: Preheat SOC medium to 37 °C
  • Transfer reaction to 1 ml SOC medium and incubate at 37 °C for at least 1 hour
  • Centrifuge 3 minutes at 12000 rpm and plate on selective LB medium
  • Incubate at 37 °C over night

2015 InterLab Protocol

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