IGEM:British Columbia/2009/Protocols/Transformation

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Transformation (adapted from OWW)

Supplies Needed:

  • Competent cells
  • Water bath/heat block at 42°C
  • Ice
  • SOC/LB medium
  • LB agar plates (with appropriate selection)
  • Plasmid DNA
  • Shaking incubator at 37°C

Steps:

  1. Remove competent cells (100uL aliquots) from -80°C and thaw on ice.
  2. Add 1uL ligation mix to thawed cells and incubate for 30 minutes on ice.
  3. Heat shock in water bath/heat block for 60 seconds and put back on ice for two minutes.
  4. Add 400uL SOC/LB medium and incubate at 37C for 1-2 hours.
  5. Plate one plate each of 100uL and 400uL, and spread evenly with sterile glass beads.
  6. Incubate overnight (<18hrs) at 37°C.
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