Long PCR

Per Mygind perm at biobase.dk
Wed Nov 12 06:12:18 EST 1997


mehboob shivji wrote:
> 
> I am having a problem amplifying a 2700 bp fragment.
> Anneal time = 2 min @ 55 deg
> Elongation = 3 min @ 72 deg
> Denature = 2 min @ 95 deg
> I would appreciate any suggestions.

Try using a much faste method, which works well in our lab.
The activity of the polymerase is inversely propotional to the time you
denature.......
Alternatively try different annealing temperatures.

Anneal time = 15 sek @ 55 deg
Elongation = 1 min @ 72 deg
Denature = 15 sek @ 95 deg


-- 
************************************************************************
If you are are not part of the solution, you are part of the precipitate
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Per Mygind, Cand.scient

Department of Medical Microbiology and Immunology
The Bartholin Building
University of Aarhus
Denmark
phone : 89 42 17 47
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