Silver Staining of DNA
dkim at nmsu.edu
Fri Dec 13 18:46:49 EST 1996
In article <58fvdu$b75$2 at mhadf.production.compuserve.com> Bob Obar <102063.2640 at CompuServe.COM> writes:
>Try calling Promega (or looking on their website). One of their DNA
>sequencing protocols relies on silver staining of single-stranded
>DNA in sequencing gels.
I have used the Silver Sequence kit for staining DNA in PAGE, but not for
sequencing purposes. I found that the protocol is easy and works
essentially as advertised, but requires development of some "hands on"
skills having to do with timing and agitation. These skills are best
learned by doing, so expect a learning curve at first.
The protocol is essentially unchanged from that of Bassam and
Caetano-Annoles' silver stain protocol which was published in Analytical
Biochemistry maybe 4 or 5 years ago. Try searching for this protocol.
If you purchase good quality chemicals (especially do not skimp on your
silver nitrate!), you can get essentially the same results with a much
cheaper home-brew kit.
My greatest problem with staining was that I would get silver deposition
on the glass plate, in spite of bind-silane attachment of the gel to the
plate. I never tried to resolve the problem, because my gels were all
about 1 or 1.5 mm thick. Instead, I just stained the gels detached from
the plate (I did lose a few gels down the drain :().
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