Taq activity at various temperatures?
jkropp at itsa.ucsf.edu
Wed May 28 17:05:05 EST 1997
> I was at a symposium of Perkin Elmer a few weeks ago and one one the speak=
> (an employee of PE) showed a graph of Taq=ADactivity vs. temperature and w=
> us that Taq has an activity at room temperature and that you should not th=
> to lightly about this.
> Cooling during pipetting is the obvious way to go and i can see the effect=
> my own PCr's and especially when i'm pipetting lots of samples. E.g. when =
> i use
> a 96=ADwell format.
Lemme respectfully suggest that there's something missing here,
viz.(whatever that means):
There's a pretty good reason for melting the ds template in the first step
of the PCR--so's the primers (hint hint) can get access to the ss target,
which the polymerase finds so sexy it ,um, jumps on and starts doing
unspeakable things with dNTPs. After which, it presumably has a cigarette.
If that primer hasn't ,er, annealed to some hot DNA our poor impotent
Mr.Taq can't "extend" anything and must content hisself with surfing the
WEB like the rest of us, looking for gifs of scantily clad inosines. Or
maybe (proof)reading Playboy...
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