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.


HTH
Dean

--
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 mailing list