Long Range PCR from yeast

Dean H. Saxe dsaxe at bimcore.emory.edu
Mon Sep 16 11:38:11 EST 1996


I am curious about the ease of doing long range PCR from yeast.  I am 
looking to amplify sequences of between 4 and 10 kb from genomic DNA.  
What I need to know are the following:

1)  Can this be done directly from colonies or do I need to first do 
genomic preps?

2)  Which kits can people recommend for performing long range PCR?  How 
reliable are the kits (i.e. how many reactions fail for unknown reasons?)

3)  I have heard that normal oligos, of approximately 20 bp, are too 
short to be used with long range PCR in other systems.  Is this true with 
yeast as well?  If so, what is the usual length for long range PCR 
primers when amplifying from yeast genomic DNA. 

Any thoughts or insights are greatly appreciated.  TIA.

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>



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