Silver Staining of DNA

[D. KIM]

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 :().

Daniel Kim