Melbourne/Transformation Protocol

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  1. Add 1uL resuspended plasmid DNA to 50uL competent cells.
  2. Incubate on ice for 45min.
  3. Heat shock in water bath at 42 degrees for 1min.
  4. Incubate on ice for 15min.
  5. Add 1mL LB.
  6. Incubate at 37degrees for 1 hour.
  7. Spin down cells and remove majority of LB.
  8. Resuspend cells in remaining LB.
  9. Under a bunsen spread resuspended bacteria on agar plate on selective antibiotic.
  10. Incubate plate overnight at 37 degrees.
  11. Place in cold room until needed.