12/07/13

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Transformation of RFP control plasmid

  • Keep competent cells on ice
    • Add 50ul of cells to 4 tubes
  • Add 1ul of resuspended DNA to 2 tubes
  • Add 1ul RFP to the other 2 tubes
  • Incubate for 30mins
  • Heat shock at 42 degrees for 60secs
  • Incubate on ice for 5mins
  • Add 200ul of SOC media
  • Incubate at 37degrees for 2hrs
  • Spread on 6 plates:
      • Positive control (normal plate)
      • Negative control (No plasmid)
      • 20ul of 5pg/ul
      • 200ul of 5pg/ul
      • 20ul of 10pg/ul
      • 200ul of 10pg/ul
  • leave plates on bench overnight