Restriction digestion of PCR fragments

Adrian Molenaar MolenaarA at agresearch.cri.nz
Thu Aug 31 16:52:58 EST 1995


In article <DDtoL0.1yL at news.cis.umn.edu>, horton at biosci.cbs.umn.edu
(Robert Horton) wrote:

> Adrian Molenaar (Molenaara at Ruakura.CRI.NZ) wrote:
> : Try two phenol extractions of your PCR product before restricting it. 
> : Taq (and sufficient nucleotides) sticks to to the DNA even
> : through ethanol precipitation in sufficient quantitites to modify your
> : newly cut ends, thus preventing succesfull ligation.
> 
> Do you have a reference for this claim? (Not that I don't believe it, I
> just can't find a reference...). Thanks.
> 
> 
> Robert M. Horton (PhD!) /\ "Crash programs fail because of the theory that
> U of M Dermatology Dept || with nine women pregnant you get a baby a month"
> Box 98 UMHC, 4-154 PWB /||\ -W. von Braun.   Disclaimer: "Bob who?"
> Minneapolis, MN 55455   ^^   horton at biosci.cbs.umn.edu       (612)625-8941

The reference is  "Cloning of PCR products can be inhibited by Taq
polymerase carryover" 1994 Biotechniques Vol 16 No.1 Pgs 32-33 by B L
Bennett and A.J. Molenaar

Regards A.M.



More information about the Methods mailing list