E.coli Transformation

•1µl of plasmid DNA was added to 50µl of competent cell (Xl-1Blue/ Stbl).

•Incubated on ice for 30 min.

•Heat shock at 45ºC for exact 90 sec.

•Incubated on ice for 2 min

•500ml of LB broth was added

•Incubated at 30ºC (Stbl)/37ºC (XL-1Blue), 200rpm for 1hr.

•Spread on LB agar plate containing appropriate antibiotic and allowed to dry.

• Incubated at 30ºC (Stbl)/37ºC(Xl-1Blue/ DH5α) for 18-20 hrs