08 August 2015 13 10K Report

Before 3 months, I had generated a clone in DH5a and subsequently in BL21 codon plus cells, that showed proper protein expression at the required molecular weight and hence had made glycerol stocks from it. Now that I am reviving it from my BL-21 stock, it is not expressing the protein at all although the clone in DH5alpha shows a proper fall out on digestion on isolating the plasmid freshly. On transforming this fresh plasmid DNA in BL21 codon plus cells, I am failing to get proper protein expression from these clones although a positive control is working which means that my experimental conditions as well as my BL-21 cells are fine. Can anyone suggest what could be the problem and where can I troubleshoot? Advice is greatly appreciated. Thanks!

Similar questions and discussions