I am working with a BAC kit and want to amplify the Red/ET and Cre expression plasmid. These two are temp sensitive and will be lost at 37C. I tried to transform them into DH5-alpha, heat shock at 42C, then incubate at 30C for 1 hour, plate onto LB with Tc, and incubate at 30C for two nights. Cre plate did not grow. Red/ET plate had a few big colonies. I inoculated 3 colonies and miniprep DNA, but got very low concentration. Anyone has similar experience? Should I incubate them at 30C longer after heat shock? Why the conc. is so low?Anyone has the sequence and map information about these two plasmid? Thanks. pls reply here or e-mail to hzs108@gmail.com