I have used SHuffle® T7 Competent E. coli from NEB and I have followed its protocol. I have used LB media and streptomycin 25 ug/mL for Shuffle cells. It seems to be important to use 30°C and not 37°C for incubation. Please, take a look on this webpage: https://www.neb.com/protocols/1/01/01/expression-using-shuffle-c3026

Vivian

I have used SHuffle® T7 Competent E. coli from NEB and I have followed its protocol. I have used LB media and streptomycin 25 ug/mL for Shuffle cells. It seems to be important to use 30°C and not 37°C for incubation. Please, take a look on this webpage: https://www.neb.com/protocols/1/01/01/expression-using-shuffle-c3026

Vivian

I agree with Vivian in following stated protocolos. In my case, cells were grown at 37 ªC and after induction, temperature was 25 ªC for 4 hours. I used LB medium with the corresponding antibiotics.

Thanks Vivian and Matias for your useful suggestions.