single colony PCR

Namjin Chung nc1 at acpub.duke.edu
Wed Sep 11 15:00:43 EST 1996


On Mon, 9 Sep 1996, your name goes here wrote:

> 
> Does anybody have a protocol for single colony PCR? (That is, use yeast
> colony directly as PCR template instead of extracted genomic DNA) And how
> well does it work? 
> 
> thank you. 
> 
> weihai (parker_lab at tikal.biosci.arizona.edu)
> 

Dear weihai,
Using pipet tips, colonies are transfered either to 10XPCR buffer and 
boiled for 3-5min or to zymolyase solution and incubated for 5min at 37C, 
and PCR'd directly.  Either way worked fine for me.  I used Pfu 
polymerase from Stratagene, but I think this method should not 
discriminate the source of polymerase.  Hope this helps, and let me know 
if you will have any problem.



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* Namjin Chung                             *                           *
* Program in Molecular Cancer Biology      * Email: nc1 at acpub.duke.edu *
* Duke University Medical Center, Box 3345 * Voice: +1 (919) 684-2363  *
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