help in PCR

Peter Ellis via methods%40net.bio.net (by pjie2 from cam.ac.uk)
Wed Jun 24 18:55:08 EST 2009


Rijuta Garapaty wrote:
> 
> Did you vary other PCR conditions, like the primer annealing temperature or
> the number of cycles? Did you add additives like BSA to check whether your
> desired product is formed? Also, did you try varying the primer
> concentration in your PCR mix? While amplifying such a big fragment, the
> choice of polymerase is also very crucial.

Since when was 1.4k a big fragment?  That should be doable with any 
polymerase on the market under standard conditions.  Check your primer 
sequence and concentration, and your DNA quality / concentration.

Peter


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