Transformation of E coli.
with Heat Shock
Thaw the appropriate amount of competent cells on ice.
Pre-chill the required number of empty 1.5 ml microcentrifuge tubes.
Pipet 50 µl aliquots of cells into the pre-chilled tubes.
Add 5-10 µl of a ligation reaction mix or 5 ng of pure plasmid DNA to each tube.
Mix gently!.
Incubate the tubes of ice for 30 min.
Heat shock the cells for 45 sec at 42°C.
Place the tubes immediately on ice for at least 2 min.
Add 800 µl of SOC medium to each tube and incubate for 1 hour at 37°C.
Transfer the cultures to 1.5 ml microcentrifuge tubes and spin for 1 min at 6000 rpm.
Remove 800 µl of the supernatant and resuspend the pellet.
Plate out the suspension on a LB agar plate containing the appropriate antibiotic.
Incubate the plates overnight at 37°C.
