native vs cloned Pfu

Wolfgang Schechinger wgschech at med.uni-tuebingen.de
Tue Sep 16 07:03:00 EST 1997


Yesterday I was talking with a Stratgene Sales Rep on this topic. He 
said that native and cloned Pfu differed in the way of purification 
and thus had different buffers. The still would sell native Pfu since 
native Pfu was first on the market and there are still people using 
it and can't or dont want to adapt their protocols set up for native 
Pfu. 
So you will have to adapt your reaction conditions to the cloned Pfu, 
or try e.g. Pwo from Boehringer Mammheim or PowerScript from PAN 
systems (also distributed at least in Germany by Biozol).

Good luck!

Wolfgang

>I was doing a PCR on some bacterial DNA with native Pfu. DNA is 
>about 64% A+T, PCR product is about 6kb, primers anneal at about 
>52.5 C. I used annealing temps from 50-52, and extensions at 72 for 
>12 min. I was getting product and pretty good yeilds. I ran out of 
>Pfu and ordered the cloned stuff from Stratagene. All of a sudden 
>the PCR isn't working so good. I notice the rx buffers are 
>different. What about efficiency, robustness etc? I'm also starting 
>to get these bright bands at about 100-200 bp. I'm basically a 
>novice at PCR so any idea on how to improve the performance will be 
>much appreciated.


>dboyd at cc.umanitoba.ca


-----                              
Wolfgang Schechinger         
University of Tuebingen
email: wgschech at med.uni-tuebingen.de
http://www.medizin.uni-tuebingen.de/~wgschech/research.htm
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