Agarose gel fluorography

Dennis Templeton djt2 at po.cwru.edu
Fri Dec 30 10:33:21 EST 1994


In article <1994Dec27.164815.1 at icbr.ifas.ufl.edu>, aris at icbr.ifas.ufl.edu wrote:

> Dear Colleagues,
> 
>         Simple question:  what is optimal method for doing
> fluorography on an agarose gel?  What is best fluor to use?

We did this recently for 3H DNA on a 1% agarose gel using the reagents
that we normally use for acrylamide fel fluorography:

1) 15 minute soak in 100% Acetic acid (to dehydrate)
2) repeat twice more
3) 45 minute soak in 15% PPO in 100% acetic acid
4) Plunge gel into cold water, wash in many changes of H20 for 90 minutes.
   (to precipitate PPO within the gel)
5) Dry using no heat for 30 minutes, then heat for 30 minutes

I recall that a colleague used to do this with 100% methanol/PPO but I
can't recall the details.

Note that while lots of folks use DMSO in the protocol above for
acrylamide gels, DMSO will dissolve the agarose. The Acetic acid left the
gel intact, and still pretty strong for handling.

Good luck,

Dennis

Dennis Templeton
Institute of Pathology
CWRU School of Medicine
djt2 at po.cwru.edu



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