Silver staining DNA

RPSCOTT RSCOTT at IRRI.CGNET.COM
Mon Aug 23 03:19:02 EST 1993


> From:	IN%"<@CGNET.COM:dkim at unm.edu>"  "dkim"    21-AUG-1993 03:43
>  
> I looked up a paper in Bio/Technology that described an arbitratrily primed
> DNA polymorphism test using the PCR.  This citation was in the Perkin-Elmer
> PCR catalog under the heading "RAPD".  I was surprised to see that the 
> detection was done by a silver staining technique instead of autoradiography.
> 
> ...Daniel Kim
> 

Silver staining affords crisper resolution of closely-spaced bands
in polyacrylamide gels than isotopic detection although sensitivity remains
comparable. 

We perform a lot of RAPD experiments here but we generally resort to
agarose-based fractionation of PCR products. Then we just ethidium-stain our
gels. We get excellent separation of 0.5-3 kb fragments on 0.50% agarose-
0.75% synergel composite in TBE.

For silver-staining the Guilemette-Lewis technique is recommended (Molec.
Genet. Anal. of Populations (1992), ed. by A.R. Hoelzel (IRL Press)):

    Guillemette, J. & Lewis, P. (1983) Electrophoresis 4:92.


Scott
======================================
R.Scott
Genetics Laboratory
Division of Plant Pathology
International Rice Research Institute 
POB 933, 1099 Manila, Philippines
e-mail: rscott at irri.cgnet.com
======================================


 



More information about the Methods mailing list