Home Protocols Gibson Assembly® Electrocompetent Cells Transformation Protocol (E5510)

Gibson Assembly® Electrocompetent Transformation Protocol (E5510)

  1. Thaw electrocompetent cells on ice.
  2. Transfer 50 μl of electrocompetent cells to a pre-chilled electroporation cuvette with 1 mM gap.
  3. Dilute assembled products 3-fold with H2O prior electroporation. This can be achieved by mixing 5 μl of assembled products with 10 μl of H2O. Add 1 μl of the diluted assembly product to 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 plates.
  10. Incubate overnight at 37°C.