Differential Display

Robert A. Anders ander at fermat.mayo.edu
Fri Apr 29 09:53:41 EST 1994


	I have a question concerning the differential display method.  I am about
to embark on this "fishing" experiment.  Another person in our laboratory
has used this method to find some interesting bands or genes already. 
These bands were rather intense and repeatable.  He subsequently subcloned
and sequenced this band from the gel.  The 120 bp segment was of no known
seq.  The segment was then used as a probe for a Northern blot.  The blot
did not show anything(controls were fine).  He has tried a larger probe and
used just poly A RNA in subsequent blots to no avail.  Here is the
question(s).  Is the absence of a good Northern reason enough to
discontinue this particular band?  Is a negative Northern common in
differential display follow up?  How about quantitative PCR(we have had a
lot of trouble with this!)?  Any thoughts on this is greatly apperciated.

Thanks in advance,
  Robert Anders.



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