Direct PCR of yeast clones picked from agar plate?

Dean H. Saxe dsaxe at
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  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:>

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