Hot start PCR

Rich Dudley rdudley+ at pitt.edu
Fri Aug 20 07:16:36 EST 1999


Really?  Thanks Duncan.  I was told Gold used a mAb.

rich

"Dr. Duncan Clark" wrote:

> In article <37BBF881.43DF51C4 at pitt.edu>, Rich Dudley <rdudley+ at pitt.edu>
> writes
> >For Amplitaq gold (PE) and Platinum Taq (Life Technologies), they use monoclonal
> >antibodies directed against the active site.
>
> Not for AmpliTaq Gold only Platinum Taq.
>
> AmpliTaq Gold is chemically modified and has a patent granted for the
> side chain modification agent they use along with claims for any other
> agents of the same chemical structure. The modification is pH sensitive
> as in reversible at low pH i.e. as you take your Tris PCR reaction
> buffer from RT to 94C it's pH drops sufficiently low enough to
> reactivate the pol. Very neat and darned difficult to find something
> that has a similar effect but is outside the patent claims.
>
> Then again it has proved quite difficult in house to force a PCR to fail
> such that hot start is needed. Spiking v.low copies of a target into
> human genomic has not reliably produced results similar to those
> published in the original antibody or oligo inhibitor papers.
>
> Duncan
> --
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
>
> Duncan Clark
> DNAmp Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
> http://www.dnamp.com
> http://www.genesys.demon.co.uk




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