A single colony was taken and inoculated overnight into 15ml LB/antibiotic at 37oC. Enough of the overnight culture was added to a basted flask the next morning, and 400-500 ml of fresh LB/antibiotic added to attain an OD600Â value of the new culture of 0. The contents were shaken for between 1 and 3 hours at 37oC until the OD600Â had reached between 0.6 and 0.8.
Upon the attainment of the correct OD600 1 ml of culture was removed and placed in an eppendorf tube labeled IPTG. The IPTG labeled tube was centrifuged for 5 minutes at 4000rpm to facilitate the draining of the supernatant. The remaining content was then stored in a freezer at -20°C to check on the gel. 1mM IPTG was added to the culture that had remained in the falcon tube and shaking continued. The entire contents of the flask were poured into the tubes after 4 hours. The tubes with the contents were centrifuged for 20 minutes at 4000rpm. The supernatant was discarded and the lysis process of the pellet continued.
