PCR to Agarose Gel Shortcuts
Fraser Dr N J
njf16154 at ggr.co.uk
Fri Oct 23 10:03:28 EST 1998
I've recently started using REDTaq from Sigma. This has the loading dye
already mixed with the enzyme and comes at the useful concn. of 1u/ul.
1ul in 25ul does the trick although 2.5/50 is recommended. You go
straight from rxn. to gel and I've had very good results with it so far.
It's reasonably priced as well.
Hope this is of some use.
Cheers - Neil.
No affiliation to Sigma etc. etc.
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.
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