Dr. Duncan Clark
drc at nospam.demon.co.uk
Mon Jul 17 03:41:00 EST 2000
In article <397196ea.2430355 at news.btx.dtag.de>, Burkhard Hassel
<Burkhard.hassel at t-online.de> writes
>your fear is unnecessary. At 37 degs the Taq-Pol is as active as a
Totally, totally incorrect.
Taq polymerase is in fact very active at 37C i.e. we find at least 10%
of the activity of at 72C. We can easily fully extend a 350bp, defined
length, primed ssDNA template in 10mins at 25C. There is more than
enough residual Taq polymerase activity left after PCR to gap fill
restriction site overhangs if any dNTPs are around.
You will have to find a PCR product purification method to be absolutely
sure of removing Taq from a PCR reaction. EtOH pptation is insufficient
(some dNTPs will also ppt with your PCR product and Taq).
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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