Q: silver stain and microsatellites

bagley at FAUNA.UCDAVIS.EDU bagley at FAUNA.UCDAVIS.EDU
Fri Jun 2 14:35:25 EST 1995


In article <1995Jun2.120202.1 at leif>, cmcgowan at kean.ucs.mun.ca writes:
>
>Helloo,
>
>I've been using microsatellite loci to do gene mapping in salmon and trout. 
>
>Presently, I end-label one of the primers with P32, run the PCR products 
>out on a sequencing gel, and then expose the gel to radiography film. 
> 
>I would like to try silver staining, however the series of 
>manipulations required is too much for the sequencing gel which is only
>0.25 mm thick and breaks apart fairly easily.
>
>Promega sells a silver sequencing kit. In it there is a reagent they 
>call 'Bind Silane' which you spread on one of your plates and it makes the 
>acrylamide gel bind hard to it. The glass plate gives the gel the support
>it needs to go through the silver staining procedure.
>
>I could buy one of these kits, but I am cheap, and would save a lot of 
>money if I could use my own silver staining reagents. 
>
>So my question is this:
>
>Does anybody know what this 'Bind Silane' is, or does anybody know of a 
>reagent that will get an acrylamide gel to bind hard (but not too hard) 
>to one of the glass plates?
>
>Thanks for your thoughts. 
>They will save me some money, and help to subvert creeping capitalism.  
>
>Colin

Sigma sells a product, methacryloxypropyltrimethoxysilane (Sigma M6514) which
works as well and may be the same thing.  Incidentally, at least with the
discount that our university receives from Promega, the cost is comparable, if
not better, when buying the kit.  Promega's protocol works better (more
sensitive and more consistent) than any others I have tried.  I would be
surprised if the cost is significantly higher than for P32 labeling when all
costs (disposal, autorad film, etc) are considered.
Mark.

Disclaimer: No relation to Promega.  Usually, not even a satisfied customer...



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