PCR on yeast colony

Kevin A. Morano kmorano at umich.edu
Mon Jun 29 08:18:08 EST 1998


In article <3596E5FF.63E8 at plovdiv.techno-link.com>,
pstoilov at obzor.bio21.acad.bg wrote:

> Hi,
> 
> I am doing this with bacterial colonies, but i think it should work with
> yeast as well. Just pick your colony with pipete tip, wash it into the
> PCR mix and run the reaction. If you are PCR-ing many colonies it is
> good idea to keep reactions on ice during colony picking.

This does indeed work, with the following caveats:

1) Use a lot of yeast, enough to make the PCR reaction turbid.

2) Use freshly grown yeast.  Cells from plates stored at 4 degrees do not
amplify well.

3)Use a little more primer (1 ug) and a little more Mg2+ (4-6 mM) than usual.

4) Use enough polymerase (1 ul Taq/tube).

Kevin Morano

-- 
Kevin A. Morano
Dept. of Biological Chemistry
University of Michigan Medical School
kmorano at umich.edu



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