Team:Calgary/Sandbox/Notebook/Protocols/BacterialTransformation

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Bacterial Transformation

Protocol

  • Thaw 100 μL of competent cells (per transformation) on ice just before they are needed;
  • Add DNA (max 20μl) thawed cells and mix by flicking the side of the tube. Leave on ice for 30 minutes;
  • Heat shock 5 minutes at 37 degrees Celsius;
  • Place on ice for 5 minutes;
  • Add 250ul SOC medium to each tube;
  • Incubate for 30 to 60 minutes with shaking at 37 degrees Celsius (note that for Kanamycin containing plasmids always use one hour);
  • Spin down to remove all supernatant except approximately 100 μL;
  • Plate approximately 50 μL on antibiotic plates;
  • Grow overnight at 37 degrees Celsius.