low plasmid yields in dh5alpha
Ed Siefker
via methods%40net.bio.net
(by ebs15242 from creighton.edu)
Wed Feb 20 13:56:30 EST 2008
Hi. We've been going through our old plasmids and making
fresh stocks. We've found a few that we're not able to
miniprep and get reasonable concentrations (<50ng/ul).
We're using Qiagen miniprep kits and invitrogen dh5alpha
and have had much success purifying
One thought was that the plasmids might be producing a
product that is toxic to dh5alpha, and maybe they'd do
better in another strain. Has anyone had a similar problem
that was resolved by growing the plasmid in another strain?
I'm thinking XL1-blue might help because it retains the
lac repressor, and that might help stop any background
expression from the lac promoter. Any other ideas would
be appreciated. Thanks
-Ed
More information about the Methods
mailing list