Gibson Assembly® Chemical Transformation Protocol (E5510)
- Thaw competent cells on ice.
- 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.
- Place the mixture on ice for 30 minutes. Do not mix.
- Heat shock at 42°C for 30 seconds. Do not mix.
- Transfer tubes to ice for 2 minutes.
- Add 950 μl of room-temperature SOC media to the tube.
- Incubate the tube at 37°C for 60 minutes. Shake vigorously (250 rpm) or rotate.
- Warm selection plates to 37°C.
- Spread 100 μl of the cells onto the selection plates. Use Amp plates for positive control sample.
- Incubate overnight at 37°C.