IGEM:Harvard/2007/Protocols/Transformation Protocol

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  1. Thaw competent cells on ice.
  2. Chill approximately 5 ng (2 μl) of the ligation mixture in a 1.5 ml microcentrifuge tube.
  3. Add 50 μl of competent cells to the DNA and mix gently by pipetting up and down.
  4. Incubate on ice for 30 minutes.
  5. Heat shock for 2 minutes at 37°C, chill on ice for 5 minutes.
  6. Add 950 μl of room temperature media and incubate at 37°C for 1 hour.
  7. Spread 100 μl onto the appropriate solid medium.
  8. Incubate overnight at 37°C.