long PCR

[peter]

On May 4, 3:53 pm, "Mandeep Sharma" <mxs... from psu.edu> wrote:
> HELLO,
>
> I am trying to amplify a 10kb maize genomic DNA fragment using long PCR.
> I have already tried Pfu ultra fusion from promega and geneamp high
> fidelity PCR system from applied biosystems and they didn't work. If you
> have tried amplification of this size fragment and you got the
> amplification then can you please suggest which taq polymerase should I
> use and what will be the most suitable conditions for long PCR with that
> taq?
>
> Thanks.
>
> Mandeep
>
> mxs... from psu.edu

When the Long PCR from genomic fails it is usually the template to
blame. I will suggest making really good high MW DNA. Recently I saw
NEB product that claims to repair template prior to PCR that might
help too. (on affiliation with NEB)
Peter