I need a good cosmid isolation protocol

Michael Kertesz kertesz at micro.biol.ethz.ch
Thu Jul 18 13:14:38 EST 1996


Hi Michael
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).
good luck
Michael


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

-- 

Michael Kertesz

ETH-Microbiology,
ETH-Zentrum/LFV,
CH-8092 Zurich, Switzerland

tel: +41-1-632 3357
fax: +41-1-632 1148
e-mail: kertesz at micro.biol.ethz.ch



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