NEBuilder® HiFi DNA Assembly Electrocompetent Transformation Protocol

For use with NEBuilder® Hifi DNA Assembly Master Mix (NEB #E2621)

  1. Thaw electrocompetent cells on ice.
  2. Transfer 50 μl of electrocompetent cells to a pre-chilled electroporation cuvette with 1 mm gap.
  3. Use 1 ul of the reaction to transform 50 ul of electrocompetent cells.
  4. Mix gently by pipetting up.
  5. Once DNA is added to the cells, electroporation can be carried out immediately. It is not necessary to incubate DNA with cells.
  6. Add 950 μl of room temperature SOC media to the cuvette immediately after electroporation.
  7. Place 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 the NEBuilder Positive Control sample. Both the transformation control pUC19 and the NEBuilder Positive Control are plated on Amp plates.
  10. Incubate overnight at 37°C.