Pfu

T. Max M. Soegaard tmms at mbio.aau.dk
Fri Oct 6 10:25:44 EST 2000


Hi.
You might also try to include betaine in your mix (final 1 M) to reduce
the difference in melting temp. between AT and GC bp´s.
Ref would be :
Hengen P, 
Trends Biochem Sci, vol 22 (1997), pp. 225-226
and cited refs therein.

Works for me on GC rich templates (genomic or cDNA).
-Max


Ali El-Salanti wrote:
> 
> Hello.
> I am having trouble with my Pfu polymerase. Template is genomic DNA and the
> size of the product should be 2 and 3 kb. With the Taq polymerase I have no
> problem amplifying the sequence, also when I try smaller bits (like 500 bp)
> I do get a product with Pfu. But I can not Pfu amplify the large sequnces.
> Maybe it should be noted that the template i 80 % AT.






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