Christopher B. Yohn
cyohn at scripps.edu
Tue Jun 6 11:03:20 EST 1995
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
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