hot-start PCR

[Marc Champagne]

In article <871719865.25332 at dejanews.com>, bqamar at paknet1.ptc.pk wrote:

> In article <33f46af4.712157 at news.unitel.co.kr>,
>   microbio at amc.ulsan.ac.kr wrote:
> >
> > Hot-started PCR was performed with paraffin wax bead.
> > It's amplification efficiency was lower than manual hot-start PCR
> > (after denaturation step, open tube cap and add Taq.pol)
> > Any thoughts about this results?
> > I think this result was due to Taq. pol degeneration during
> > degeneration step or uncomplete melting of wax-layer..
> > but I can't sure..
> 
> If you want to do hot start reactions I will recommend TaqGold from
> PE/ABI the TaqStart Antibody from Clontech.  I have used both of them and
> find them quite satisfactory.  Get rid of the Wax once and for all.
> 
> Raheel Qamar
> 
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Do you really need hotstart?  I've found that with most primer sets i can
decrease my spurious bands by simply mixing my reactions at room temperature,
instead of on ice, and by starting each PCR run with a 2 min at 94 extra-step.

m

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