Long PCR

Chang Zhu czhu at changbioscience.com
Tue Apr 6 19:00:48 EST 2004


Hi,

I've been attempting to develop a PCR primer design program for
difficult template. Could you give a try? The program is at the
following link:

http://www.changbioscience.com/primo/primohl.html

I'll appreciate your feedback. Thanks.

Chang

ashiflett at mbl.edu (AShiflett) wrote in message news:<1b3ed0cb.0403251313.9c917f1 at posting.google.com>...
> Hi!  I am attempting to amplify an approximately 12 kb piece of DNA
> that is near the telomere of a chromosome.  I have used both Herculase
> from Promega and the Elongase enzyme from Invitrogen.  In both cases,
> I do not obtain a product of the expected size.  I followed the mfg
> directions, and performed optimization for primers, annealing temp,
> and Mg concentration.  I use approximately 100 ng of genomic DNA which
> is not sheared and looks great on a gel.  Does anyone have any tips
> for amplifying long pieces or DNA, or can recommend a good polymerase
> that has been successful for difficult PCRs?  Any advice is welcome
> and appreciated.



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