PCR to Agarose Gel Shortcuts

Carl Simard c.simard at sympatico.ca
Thu Oct 22 17:15:44 EST 1998


If you don't have to manipulate the amplified DNA after the PCR, you can put
the loading buffer directly in the product tube (after the amplification is
done!) and load what you want. I've done that many time and there is no
problem. The only thing you should care is to wipe the mineral oil that
stick on the exterior surface of the tips you use to load.

Carl Simard
c.simard at sympatico.ca

John Scott Meschke wrote:

> Does anyone having time saving shortcuts for running PCR products on an
> agarose gel?
> My current protocol involves aliquoting 15 microliters (of 100) of
> product with 3 microliters of 1x gibco loading buffer.  I am considering
> skipping the aliquot step and putting loading buffer directly into my
> product tube.  This will eliminate a centrifuge step, and at least one
> pippetting step.  What I am concerned about is the stability of the cDNA
> in the presence of loading buffer over time.  The assays I am running
> are merely presence absence and I do not need to manipulate the DNA
> after running the gel except for gene probing for confirmation. Does
> this sound like a good idea?  Are there any other time savers out
> there?  I have considered red taq, but I am concerned with the potential
> differences between the Amplitaq that I currently use and the red taq.
> My experiments are long term comparisons and I hate to modify my
> protocols too much over the course of the experiment.  Thanks alot.
>
> Scott




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