SDS in PCR?

tweissen at my-deja.com tweissen at my-deja.com
Wed Jan 5 07:32:20 EST 2000


'Happiness' of the enzyme is perhaps not the most
important factor.
Some additives reduce Taq activity but can still
increase yield by improving template denaturation
in each cycle - depending on how well the
template denatures normally and whether or not
an excess of enzyme is used.

Thomas Weissensteiner

Edward Jenner Institute
for Vaccine Research
Compton RG20 7NN, UK


In article <9PqRmWAEugc4EAOj at genesys.demon.co.uk>,
  "Dr. Duncan Clark" <Duncan at genesys.demon.co.uk>
wrote:

>
> For Taq polymerase
>
> Concn (presume final)        Activity
>
> 0                               100%
> 0.001%                          105%
> 0.01%                           10%
> 0.1%                            <0.1%
>
> However addition of 0.5% each Tween20 + NP40
reverses the inhibitory
> effect of 0.01% SDS.
>
> The enzyme is also happy in 2M Urea, 10%
ethanol, <5% DMF and <10%
> formamide.
>
> All from PCR Technology, Principles and
Applications for DNA
> Amplification, Ed. Henry Erlich, Stockton
Press. Table 1, page 20 in the
> chapter written by David Gelfand. Of course
Promega may dispute this
> information. I merely pass it on without too
much comment.
>
> 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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