Help: high mutation rate during RT-PCR
Dr. Duncan Clark
Duncan at nospam.demon.co.uk
Tue Aug 15 11:53:46 EST 2000
In article <8nbe8q$rk7$1 at uni00nw.unity.ncsu.edu>, Karthik Aghoram
<kaghoram at unity.ncsu.edu> writes
>I don't think Advantaq has proofreading activity
It hasn't. I'd say it is basically a Klentaq.
>Pfu has proofreading activity. But it is not as efficient as Taq, and
>you may have to double your extension periods.
Can I just qualify this for other readers in case it is mis-interpreted?
Karthik is referring to the polymerase efficiency as a whole and not the
efficiency of the proof-reading activity. Taq has no proofreading
>Roche Boehringer has an "Expand Hi-fidelity" system that has both Taq and
>a profreader (Pow?). It has worked well for my colleagues here.
It has a reported fidelity of roughly one third of Pwo or Pfu on their
own. Likewise the mix has a three fold or so better fidelity than Taq.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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