PCR can not get intense bands

Bernard Murray bernard at elsie.nci.nih.gov
Fri Mar 22 13:48:19 EST 1996


In article <Pine.SUN.3.91.960322100522.16700A-100000 at verdi>, hroychow at NMSU.EDU 
says...
>
>On Fri, 22 Mar 1996, Zahra Fayazi wrote:
>
>> Hello everyone
>> I am trying to amplyfy a 421bp region
>> I can only get a very faint band.

Assuming reasonably clean DNA, reasonably well chosen primers and
reasonably optimised Mg and temperature the "magic fix" I've
loved for strong amplification of genomic DNA is the addition of
DMSO to 5%.  This has worked wonders for me.  Increasing the
amount of template certainly has to be done carefully as you can
only go so far before you see inhibition.  I have also heard people
rave about some of the new enzyme cocktails.  The most popular in
this neck of the woods is Boehringer's "Expand High Fidelity"
which seems to give better amplification of small bands than Taq alone.

		Good luck,
			Bernard

Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov  (National Cancer Institute, NIH, Bethesda MD, USA)




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