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Reamplify PCR product

Mark S. Strom strom at listeria.nwfsc.noaa.gov
Tue Dec 5 16:50:31 EST 1995


Dirk Mueller wrote:
> 
> I have a lot of trouble with reamplifying my 1150bp PCR Product.
> I tried the reamplification with diluted PCR product with and without
> purification with Qiaquick coloumn. The result is nearly the same. Only 6%
> DMSO gave some tolerable results.

I've had good luck doing a second PCR on a fragment after running it through an 
agarose gel--the procedure (first posted in this newsgroup by Ed Rybicki back in 1992) 
involves taking a small "core" or plug of agarose containing the band and reamplifying 
that directly.  The procedure is spelled out at 
<http://research.nwfsc.noaa.gov/protocols/core_sample_PCR.html>.

Good luck!

-- 
Mark S. Strom, Ph.D.                 (206) 860-3379  
Utilization Research Div.            (206) 860-3394 (Fax)
Northwest Fisheries Science Ctr.     mstrom at sci.nwfsc.noaa.gov   
NMFS/NOAA                            strom at listeria.nwfsc.noaa.gov
2725 Montlake Blvd. E.              
Seattle, WA  98112-2097
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