plasmid isolation from yeast

Namjin Chung nc1 at acpub.duke.edu
Fri Sep 13 08:27:48 EST 1996


On Wed, 11 Sep 1996, Kevin A. Morano wrote:
> 
> 
> "Rapid isolation of plasmid DNA from yeast",  Current Protocols,
> contributed by Charlie Hoffman
> 
> Breaking buffer:  2% (v/v) Triton X-100, 1% (v/v) SDS, 100 mM NaCl, 10mm
> Tris 8.0, 1 mM EDTA
> 
> It's a great protocol!
> Kevin
> 
> -- 
> Kevin Morano, Thiele Lab
> University of Michigan Medical School
> kmorano at umich.edu
> 
> 

I have also wondered if there is any method like that for a long time, 
and I check the red book.  As I see it, it was not so new method, neither 
it was not for 'isolating' plamid directly from yeast cells without 
transforming bacteria.

The method is still considered to be a good one, and it is to prepare 
yeast lysates which contain plasmids in addition to chromosomes for 
bacteria transformation.  Since yeasts are eukaryotes, both DNA types 
will be released into lysates when you break the cells.  Then, I guess it 
is not easy to separate plasmids from chromosomes.

As I said, however, it is still fast and good method to isolate 'total' 
yeast DNAs for bacterial transformation.  


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* Namjin Chung                             *                           *
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