I need a good cosmid isolation protocol
kertesz at micro.biol.ethz.ch
Thu Jul 18 13:14:38 EST 1996
Retry the Qiagen kit. If you grow a 500 mL to 1 litre LB culture of the
strain with your cosmid, and then do a normal alkaline lysis very, very
gently (use a big centrifuge bottle for the neutralization step, and just
it rock it to and fro a few times to mix), and then apply it in two or
three goes to the Qiagen column (reequilibrate each time with QBT), it
should work ok, and give you a clean prep, and decent yield. Don't bother
with smaller volumes - it's not worth the effort (I had the same trouble
as you six months back).
Michael R. King wrote:
> Can anyone give me a reference to a protocol giving high yield cosmid
> isolation. I've used the kits like Qiagen but they don't work very well.
> I'm doing a basic mini-prep as I type this but I thought I would get a
> little advice.
> Kind regards,
> Michael R. King
CH-8092 Zurich, Switzerland
tel: +41-1-632 3357
fax: +41-1-632 1148
e-mail: kertesz at micro.biol.ethz.ch
More information about the Methods