Gibson Assembly® Chemical Transformation Protocol (E5510)

  1. Thaw competent cells on ice.
  2. Add 2 μl of the chilled assembly product to the competent cells. Mix gently by pipetting up and down or by flicking the tube 4–5 times. Do not vortex.
  3. Place the mixture on ice for 30 minutes. Do not mix.
  4. Heat shock at 42°C for 30 seconds. Do not mix.
  5. Transfer tubes to ice for 2 minutes.
  6. Add 950 μl of room-temperature SOC media to the tube.
  7. Incubate the tube at 37°C for 60 minutes. Shake vigorously (250 rpm) or rotate.
  8. Warm selection plates to 37°C.
  9. Spread 100 μl of the cells onto the selection plates. Use Amp plates for positive control sample.
  10. Incubate overnight at 37°C.