SSCP help

Frederick Garbrecht FRED at bmt.mcw.edu
Wed Sep 7 14:40:28 EST 1994


I am trying to do a single stranded conformational polymorphism 
analysis on a ~500 bp PCR fragment.  I run the formamide denatured 
fragments out alongside non-denatured control fragments on a 4% PA 
gel and then silver stain.  The non-denatured fragments give nice 
sharp bands, but the denatured fragments give a fairly broad smear, 
in which I cannot discern individual bands.  I assume that the 
smearing has something to do with the denaturation (the bands appear 
pretty high up on the gel, which is where they ought to be).  I am 
using deionized formamide.  Any hints from the experts on how to turn 
my smears into focused bands?  Thanks alot--

Frederick C. Garbrecht
fred at bmt.mcw.edu
fgarbrec at post.its.mcw.edu
Bone Marrow Transplant Program
Medical College of Wisconsin
phone 414 257 5053
fax   414 257 7994



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