Differential Display of message tips?

Randy Hardy hardy at mighty.fccc.edu
Mon Nov 29 15:02:58 EST 1993


 I wonder if anyone out there has experience with the differential display of
message technique.  We are in the initial stages of setting it up (made 3 5'
and 3 3' oligos) and have had variable success so far.  When it works we see
reasonable bands with an overnight exposure of the gel.  However, we often
simply get faint smears of signal all down the lanes (no discrete bands).  As
far as we can tell this is all with good cDNA (specific test oligos give very
good bands).  Titering amounts of Mg and oligos and cDNA does not appear to
help much (already optimized).  Any pointers on what we should look out
for/check?  One suggestion was that the radiolabeled ATP was incompletely
thawed/mixed, although this is not established as the problem.  Any advice is
appreciated.



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