IGEM:British Columbia/Protocols/Transformation Competent Cells

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Materials

  • Competent cells
  • Plasmid to be transformed
  • Ice
  • LB medium
  • LB agar plates

Equipment

  • Water bath/heat block at 42°C

Method

  • Remove 100μL aliquots of competent cells from -80°C freezer and thaw on ice
  • Add 1μL of plasmid to be transformed to the thawed cells and incubate for 30 minutes on ice

NOTE: If working from a miniprep plasmid stock, the concentration will likely be too high and will have to be diluted.

  • Heat shock in water bath/heat block for 60 seconds
  • Put cells back on ice for 2 minutes
  • Add 400μL LB medium and incubate at 37°C for 2 hours
  • Plate entire 500μL onto a LB agar plate with the correct antibiotic and spread evenly with a sterile spreader
  • Incubate overnight (<18hrs) at 37°C
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