Promega Silver Sequencing

[Daniel Kim]

RE:  smeared bands in Promega silver sequence system.

I haven't used this system, but have used silver stain of DNA for DDRT 
gels.  I have found that the silver stain does not have any effect on the 
banding.  You might be having trouble related to the bind silane that 
keeps the gel attached to one of the glass plates.

You might consider running a few radiolabeled sequencing reactions on the 
same gel and doing an autoradiogram of the stained gel, to see if bands 
show up that way.  If so, your problem may relate to the fact that the 
silver stain is nonspecific (that is, you may have extraneous DNA in your 
reaction mixes that are not labeled by the DNA polymerase, but are picked 
up by the silver.).

Daniel Kim
dkim at nmsu.edu