Are you sure it is not a phage infection that is causing lysis of your cells?
Christopher J. Law, PhD Skirball Institute of Biomolecular Medicine, Structural Biology (3-5), New York University Medical Center, 540 First Avenue, New York, NY 10016, USA. 2008/4/30 Raji Edayathumangalam <[EMAIL PROTECTED]>: > Hi Folks, > > I am working with E. coli cells co-transformed with two plasmids and I find > that my cells lyse > following overnight inductions at 18C. I suspect (among many things) that > Ampicillin+ > Chloramphenicol+ Kanamycin in the medium may be the source of my woes. > > My colleagues have suggested growing cultures at 18C, say for 4-6h instead. > Has anyone had > reasonable protein expression levels by inducing cultures at 18C for 6h? > From what I understand, the > E. coli doubling time is manyfold longer than at 37C. But I thought I'd ask. > > I am already playing with lowering and/or doing away with the antibiotics. > > Any suggestions wrt 18C? The protein is insoluble at 30C. > > Thanks. > Raji >
