>I am trying to do PCR from yeast plasmids and am not having any success.
>I have made 'yeast extract' according to the protocol in the Red Book
>(current protocols in molecular biology), I can transform E. Coli with the
>extract, but for some reason my PCR is not working. Is there something
>that I am missing? Any help would be greatly appreciated, Thank You
There is an extensive discussion of how to PCR from yeast colonies in the
archives of this listserve, accesible through the Saccharomyces Genomics
Database Web page (http://genome-www.stanford.edu/). In brief, we do the
1. Take 1/4 to 1/2 of a freshly grown yeast colony on the end of a clean
toothpick. Resuspend in a conventional 50 ul PCR reaction.
2. Do a standard PCR.
In other words, there's no need even to purify your plasmid.
lichten at helix.nih.gov