Cosmid purification

[Christopher B. Yohn]

In article <3qo0mg$aol at dartvax.dartmouth.edu>,
Mari.L.Shinohara at dartmouth.edu (Mari L. Shinohara) wrote:

> Hi netters,
> I'm having a hard time purifying a cosmid clone.  
> I understand that handling cosmids could be sometimes tough.  I always
> streak clonies one day before purification, then Terrific Broth was
> used for bacterial propagation.  I've got a small amount of cosmid with
> Qiagen only once out of six trials:  Usually, I didn't get any. 
> (Miniprep seems to work, but yield is terribly low.)
> 
> I would greatly appreciate if anyone can give suggestions.
> 
> Mari Shinohara
> Dartmouth Medical School

I found that the method you use for your cosmid miniprep can affect your
yield.  Using the Merlin (diatomaceous earth) or LiCl methods gives low
yield, while using a standard alkaline lysis miniprep protocol gives
better yields (in my hands).  Try different miniprep protocols until you
find one that works.

-- 
Christopher B. Yohn  [cyohn at scripps.edu]
The Scripps Research Institute
Dept. of Cell Biology - MB8
La Jolla, CA  92037
Ph: (619) 554-4335
Fax: (619) 554-6188