Dear friends, I am sorry for off-topic though it may be related indirectly! I mutated a 60kDa protein (changing from X-->Pro). After doing all the steps, I sequenced the expression vector. It seems everything is fine, even with promoter, but, I can't express it. I tried different Tempratures, IPTG (with new Stock), media, but they didnot make any difference. Any suggestion is highly appreciated.
Best, Jahan
