Isolation of Yeast/E. coli Vectors

tronni at my-deja.com tronni at my-deja.com
Sat Nov 18 19:06:48 EST 2000


In article <3A15AB2E.11FA6F53 at rhrk.uni-kl.de>,
  =?iso-8859-1?Q?J=F6rn?= Jeblick <jeblick at rhrk.uni-kl.de> wrote:
> Hi there!
> Is there any way to isolate a Yeast/E. coli shuttle-vector from S.
> cerevisiae without getting all the (sheared) genomic DNA, too?
> Thanx a lot!
> Jörn Jeblick
>
>

There is a method for yeast plasmid isolation in the
"Current Methods in Molecular Biology", it's based on
glass beads + breaking buffer + phenol-chloroform. Cheap & works!
The resulting aqueous phase contains plasmids and sheared genomic DNA
but this does not matter for next step. Yield is always poor from yeast.
You can rescue your plasmid by transforming highly competent E. coli
(transformation efficiency has to be at least 1 x 10E7/ug) and growing
up the clones.

Best regards,

Tapani Ronni, Ph.D.
HHMI/UCLA


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