Transformation

Use up to 5 μL of the ligation mixture or up to 10 ng of supercoiled plasmid for the transformation of 50 μL of chemically competent E. coli cells. If cells were frozen thaw them on ice.

Add up to 1/10 volume of DNA and incubate on ice for 30 minutes.
Heat-shock cells at 42 °C for 1 minute and transfer cells back on ice.
Immediately add 1 ml of LB medium and incubate at 37 °C for 1 hour.
Centrifuge and resuspend cells in a low volume of LB medium.
Plate on LB-agar plate supplemented with an appropriate antibiotic.
Incubate overnight at 37 °C.