Silver sequencing

Jerry Kropp jkropp at itsa.ucsf.edu
Thu Oct 17 21:19:23 EST 1996


In article <pezmen.1.00099753 at gate.net>, pezmen at gate.net (Hector
Cruz-Lopez) wrote:

> anybody using this protocol?  we just lost our radioactve materials
license to 
> budget cuts and need an alternative.  all responses will be appreciated - we 
> are desperate!
> 
> hector cruz-lopez
> fisheries genetics research lab

I haven't tried silver, but routinely seq PCR fragments of human genomic
DNA using a biotinylated seq primer and the NEB Phototope kit for
detection. The bands arent as sharp as with S35, due to the fact that a
transfer of the seq gel (quite easy) causes some diffusion. However the up
side is I expose the film for 10 min.
It also works with cycle seq with Thermosequenase. If you want I'll send
you the protocol, but I'll be out of town (diving in Mex--eat yer heart
out) from 10/18 till 10/28.
Good Luck,
Jerry



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