Troubles with Pfu polymerase

[Holger Scholz]

In article <4cjekp$ll0 at saphir.jouy.inra.fr>, hatzfeld at msdos.ensam.inra.fr
(Yves hatzfeld) wrote:

> Hi Netters!
> 
> I have troubles with amplifying a 1.5 kb fragment with pfu polymerase
> (stratagene). 
> 
> When I do the same experiments using Taq instead, I haven't any problem
> and I obtain the expected products ...
> 
> So why the Hell it does not work?
> 
> Any idea would be very wellcome, before I encounter a nervous
> breakdown...

> 
> Yves.

It´s always the same story with pfu polymerase.

try 100 microliter reaction volume not 50
increase the primer concentration, 500 ng each

I don´t know why but it is like it is
   
                   holger