Growing E.Coli at Different Temperaturesdo - (Jan/13/2009 )

Hi,

I am very new to biology and have a basic question about growing E. Coli.

I want to overexpress proteins from E. Coli at different temperatures (maybe about 16C, room, 37C). When I grow the overnights do I grow them all at 37C and then dilute and induce expression at the particular temperature or do I grow the overnights at the temperature of dilution and induction?

Your help is very much appreciated.

What I have always done is grow the overnights at 37, dilute the cultures and grow those at 37 until they reach the desired OD for induction, then cool the culture on ice for 45 min before inducing at the desired temperature. This cold shock seems to help a little bit, at least with preventing the formation of inclusion bodies when we want to do a native purification and not a denaturing prep.