"Pfx" DNA pol...

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Fri Jun 22 04:57:02 EST 2001


Duncan, 

think of any reaction that reversibly might modify an enzyme 
and may be perfomed at RT without pH change and catalyts. 
Reversion should be temperature dependent i.e. no chemicals 
than those already present in PCR mix should be necessary..

In the patent, they suggest formation of a Schiff base (-NH2 + 
+HCHO <-> -N=CH2 + H2O). Unless you reduce that stuff 
(textbooks use borohydride), just add lots of water and and the 
imine will revert. The funny thing however is that Q claims the 
modification even being stable after removing excess 
formaldehyde. 

But maye someone just simply did in situ PCR on formaline 
fixed thermophile sp. and forgot to add polymerase but got 
results...

Wo

> But back to science ....
> 
> How on earth did Qiagen come up with the idea that by treating
> thermophilic enzymes with formaldehyde they would be able to recover
> activity after heating them?  
> 
> Luck? 
> 
> Accident?
> 
> I know they quote some prior art that RNase survives formaldehyde but it
> is a big jump from that to a HotStart polymerase.
> 
> All credit to them.
> 
> Duncan   
> -- 
> The problem with the gene pool is that there is no lifeguard.
> 
> Duncan Clark
> GeneSys Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
> 
> 


[tiss mezzage wahs broduceRd using TYPO GENERATOR zoffwer]
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Dr. Wolfgang Schechinger
Lab N233 (c/o Dr. Steve Roffler)
Institute of Biomedical Sciences
Academia Sinica
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Tel +886-2-2789-9152
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