Direct PCR of yeast clones picked from agar plate?
Dean H. Saxe
dsaxe at bimcore.emory.edu
Tue May 7 13:56:02 EST 1996
I have good luck using the my standard PCR reaction conditions (200uM
dNTPs, 1X Taq buffer, .75 pg each primer, 1 - 2 units Taq) but I
double the MgCl2 concentration to 3 uM (otherwise I use 1.5 uM for
normal PCR reactions). I smear a small amount of a colony on the
bottom of the PCR tube, add my reagents and mineral oil and run the
reactions as normal. The first cycle at 94 degree C bursts enough of
the yeast cells to provide a starting template for amplification.
Dean H. Saxe "Life gives us hurdles, and we must jump,
dsaxe at bimcore.emory.edu and if you fall, you get your ass back up!"
-Millan & Kenzie "Blow"
Graduate Student, Dept. of Genetics & Molecular Biology, Emory Univ.
My home page --- <URL: http://userwww.service.emory.edu/~dsaxe>
More information about the Yeast