hot-start PCR

Harry Witchel Harry.Witchel at Bristol.ac.Uk
Tue Aug 19 17:11:54 EST 1997


Hi --
	You do not mention what size tube you were using with your wax 
beads.  If you use smaller volumes than the manufacturer recommends, the 
reaction will be inefficient because the two phases will not mix 
properly after the wax melts.  Normally, the fluids mix by convection.
	Harry

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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