Silver staining of aDNA in agarose gels

Philip L. Carl plc at med.unc.edu
Thu Apr 17 09:38:44 EST 1997


We are initiating some differential display experiments, and as several 
sources have suggested that the traditional denaturing acrylamide gels in fact 
yield many "artifactual" bands I have been doing some preliminary experiments 
using FMC Metaphor agarose for displaying the bands.  The gels are much 
stronger than low concentration acrylamide gels, and I suspect that another 
advantage of agarose is better recovery of bands for reamplification.  The 
problem is detecting the bands.  I'd like to avoid radioactivity if possible 
for the usual reasons.  We have tried SYBR Green I, but like some others find 
that it is not very much more sensitive than EB-perhaps as much as a factor of 
10 fold (i.e. a few ng), but probably less in our hands-certainly not the 60 
pg claimed in the tech literature.  We've also tried silver staining using the 
method of Gottlieb and Chavko Anal. Biochem. 165: 33-37 (1987) which  claims a 
lower limit of detection of 2.5 ng-a claim we believe reasonable.  Trouble is 
that this is probably about 1/100 as sensitive as staining acrylamdide with 
silver.  So my question is, does anyone know of a better silver stain for 
agarose gels? 

Philip L. Carl
Research Associate Professor of Pharmacology and
Associate Director The Program in Molecular Biology and
Biotechnology
101 MBBRL CB7100
Medical School
University of North Carolina at Chapel Hill
Chapel Hill, NC 27599-7100
Phone: 919-966-3544
FAX: 919-966-6821
email: plc at med.unc.edu



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