Re-transformation of DH5-alpha cells with plasmid DNA

"Heat Shock"

  • Thaw out the competent cells on ice
  • Put plasmid DNA (0.1 ng) in a Eppi tube
  • Add 50 –100uL competent cell to DNA, mix gently and leave in ice.
  • Heat shock at 42 ° C for 45 –120 sec (use water bath or heat block)
  • Incubate in ice for 2 mins
  • Add 300uL SOC medium to cell and mix
  • Incubate at 37 ° C for 15 mins
  • Spread with glass spreader on Solid LB medium
  • Incubate at 37 ° C overnight.