PCR - Master Mix

John Scott Meschke jmeschke at sph.unc.edu
Tue Nov 17 10:42:23 EST 1998


I second the suggestion for making a large batch of master mix.  However, I
suggest aliquoting your mastermix into your reaction tubes and then freezing
them down, rather than freezing the bulk down and aliquoting as needed.  If you
are worried about evaporation there are PCR compatible waxes or parafin that can
seal your mix in the reaction tube.  I believe there was a Biotechniques article
about 6 months to a year ago which compared reaction quality over time using a
similar method.  I have used this method for the last 6 months, without any
detrimental effects to my PCR or RT-PCR.  My normal storage time is generally
less than a month and I make up roughly 110 reactions at a time.  This method
also allows standardization of the reaction mix for long term or replicate
experiments.

Caddie wrote:

> Simply make your own master mix by mixing a large prep of buffer, water,
> MgCl2, and primer(s).  Then when you are ready to PCR, aliquot out what you
> need, mix in the appropriate polymerase, and add to your samples.
>
> Caddie
>
> <><><><><><><><><><><><><><><><><><><><><><><>
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>
> beakerhead at hotmail.com wrote in message <72n9st$otv$1 at nnrp1.dejanews.com>...
> >Hi!  I'm looking for a master mix for my PCR reactions. I'd love to just
> put
> >in my template and primers and not have to do all the pipetting of other
> >reagents.  Does anyone have any experience with any products out there?
> >Please send me any info-experiences that you might have.
> >
> >Thanks
> >beakerhead at hotmail.com
> >
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