hot-start PCR

bqamar at paknet1.ptc.pk bqamar at paknet1.ptc.pk
Wed Aug 20 22:51:14 EST 1997


In article <M.Champagne-1908971208070001 at ocarina.cellbio.duke.edu>,
  M.Champagne at nospam.cellbio.duke.edu (Marc Champagne) wrote:

> > If you want to do hot start reactions I will recommend TaqGold from
> > PE/ABI or 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
> >
> > -------------------==== Posted via Deja News ====-----------------------
> >       http://www.dejanews.com/     Search, Read, Post to Usenet
>
> 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.
>
> Pls remove -nospam- from my address if you reply by e-mail

The reason that I use the TaqStart antibody is that I am multiplexing at
least six different microsatellites in one tube (i.e. multiplexing the
reaction and not pooling afterwards like traditionally it has been done)
and it really helps in eliminating spurious bands.  BTW I am using
fluorescent primers and the ABI 377 to separate and size my fragments.

Multiplexing saves not only on the labor but also the amount of primer and
enzyme that you have to put in each reaction.  Since you can get away with
having a lot less of these two in each tube.

Try Multiplexing and you won't ever go back to single reaction per tube.

Raheel Qamar

-------------------==== Posted via Deja News ====-----------------------
      http://www.dejanews.com/     Search, Read, Post to Usenet



More information about the Methods mailing list