low plasmid yields in dh5alpha
(by take_murata from hotmail.com)
Fri Feb 22 07:59:41 EST 2008
Reduce the cultivation period. During long period, E. coli may lose plasmid but get anti-antibiotics secreted in media and E. coli can be alive.
> Hi. We've been going through our old plasmids and makingfresh stocks. We've found a few that we're not able tominiprep and get reasonable concentrations (<50ng/ul).We're using Qiagen miniprep kits and invitrogen dh5alphaand have had much success purifying>One thought was that the plasmids might be producing aproduct that is toxic to dh5alpha, and maybe they'd dobetter in another strain. Has anyone had a similar problemthat was resolved by growing the plasmid in another strain?>I'm thinking XL1-blue might help because it retains thelac repressor, and that might help stop any backgroundexpression from the lac promoter. Any other ideas wouldbe appreciated. Thanks>-Ed
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