Gibson Assembly® Electrocompetent Transformation Protocol (E2611)
- Thaw electrocompetent cells on ice.
- Transfer 50 μl of electrocompetent cells to a pre-chilled electroporation
cuvette with 1 mM gap.
- 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
- 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
- Place the tube at 37°C for 60 minutes. Shake vigorously (250 rpm) or
- Warm selection plates to 37°C.
- Spread 100 μl of the cells onto the plates.
- Incubate overnight at 37°C.