Query on silver staining of PAGE/Hydrolink gels
jbock at HOBBES.KZOO.EDU
Wed Oct 27 09:19:41 EST 1993
Jacquie Greenberg wrote:
> Dear Paul,
> I received the letter below from our local biotech news and now have
> a query which I believe would be appropriate to channel through you.
> I am presently involved with mutation screening using SSCP analysis
> and the method of visualising the results that I have chosen is to
> silver stain the hydrolink gels. I prepare my plates so that the gel
> will stick to one plate when I serepate them after electrphoresis
> and then I stain it on the plate (the "glue" I use is MTMS and
> glacial HOAc in ethanol)
> The problem now comes when I want to remove the gel once it is
> stained and transfer it filterpaper to dry it, it will not come off!
> I have tried soaking it in 2% NAOH, but then it floats off, stretched
> out of all proportion and the results are distorted.
> Is there any secret method to release it successfully from the
> plate? I really would appreciate some advise in this regard.
> Many thanks for your help. If you are NOT the right person to
> contact, please could you let me know how to go about getting this
I assume that MTMS is gamma-methacryloxypropyltrimethoxysilane. The
purpose of the "glue" is to keep the gel in place on the glass support
during the staining process _and_ to eliminate the need to transfer the
gel to filter paper to dry. The gel is dried to the plate either by
air drying overnight, or in a 37C room (oven, etc.) for a few hours.
When the gel is totally dry it is stable until purposely removed by the
NaOH (or 2% KOH, which does not etch the glass as readily). This allows you to
take a picture of the gel for a permanent record, or expose to X-ray film,
if necessary, by placing the entire glass plate into the exposure
I hope this helps. I have used this "gel-goo" process for sequencing gels for
years and would not go back to transfering the gel to filter
Joyce V. Bock
Kalamazoo College Biology Dept.
Kalamazoo, MI 49006-3295
jbock at kzoo.edu
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