NEBuilder® HiFi DNA Assembly Electrocompetent Transformation Protocol
For use with NEBuilder® Hifi DNA Assembly Cloning Kit (NEB #E5520)
- Thaw electrocompetent cells on ice.
- Transfer 50 μl of electrocompetent cells to a pre-chilled electroporation cuvette with 1 mM gap.
- Use 1 ul of the reaction to transform 50 ul of electrocompetent cells.
- Mix gently by pipetting up.
- Once DNA is added to the cells, electroporation can be carried out immediately. It is not necessary to incubate DNA with cells.
- Add 950 μl of room temperature SOC media to the cuvette immediately after electroporation.
- Place 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 plates.
- Incubate overnight at 37°C.
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