PCR from genomic DNA

Stephen R. Lasky Stephen_Lasky at brown.edu
Fri Apr 15 07:36:25 EST 1994


In article <1994Apr14.200642.2256 at alw.nih.gov>, Jim Owens
<jow at helix.nih.gov> wrote:

> In article <199404061949.AA00341 at wugate.wustl.edu> ,
> krishnan at BORCIM.WUSTL.EDU writes:
> >Could somebody tell me how to PCR-amplify a 3 kb piece of DNA from total
> human
> >genomic DNA or from total yeast DNA
> 
> In addition to what others have said:
> 
> If your DNA is very viscous, you should consider making it less viscous
> by some vigorous vortex mixing or by digesting with a rare cutting
> restriction enzyme that does not cut between your PCR primers.
> 
> Good luck,
> 
> Jim Owens


In my experience, if your DNA is viscous, you have too much for PCR. 
Dilute it at least 1000 fold, mix, and then use one microliter of that for
PCR.  
-- 
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Stephen R. Lasky, Ph.D.       Brown University/Roger Williams Medical
Center
e-mail: Stephen_Lasky at brown.edu         LandLine: 401-456-6572
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A nuclear war could ruin your whole day.
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