In article <4m6t2p$5nu at seagoon.newcastle.edu.au>,
<@medicine-dmb.newcastle.edu.au> wrote:
> I'm having trouble PCR-ing from my pGAD10 Two-Hybrid activation domain
> vector miniprepped from yeast (lots of non-specific bands and absence of
> bands in some preps). Has anyone got any suggestions for PCR-ing from
> yeast minipreps, or does it depend mainly on the primers used? Also does
> anyone know how quickly yeast lose plasmids? Is it OK to grow up yeast
> for a plasmid prep in rich media, or is it better to use selective
> media?
>> Thanks,
> Estelle
------------------
Hi Estelle
Just use your colony on the plate for PCR. After suspending
the colony, you can start the reaction. Or, clean up your template by using a
resin from some companyUs minipreps kit.
Hope this helps
Nobuo SAKATA, Ph.D.
Showa Coll Parmaceut Sci
