• Thaw all reagents completely on ice.
  • Add 1 µL of ligation reaction to thawed competent cells.
  • Gently mix by tapping tube of competent cells.
  • Incubate reaction on ice for 30 minutes.
  • Heat shock the competent cell mixture by incubation for 30 to 60 seconds in a 42°C water bath.
  • Incubate tubes on ice for 2 minutes.
  • Add 250 to 500 µL of SOC or LB media.
  • Incubate at 37°C and shake at 250 rpm.
  • Warm selection plates to 37°C.
  • Spread  10, 50, and 100 µL of transformed cells on  selection plates.
  • Incubate plates at 30°C overnight.
Component
Final concentration/amount
Competent cells
to 50 µL
Ligation reaction
1-5 µL
SOC or LB media
950 ml

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