"reverse northern" problems

Ed Wang ed_wang at arris.com
Fri Dec 6 09:54:15 EST 1996


Hello,
Let me introduce myself as a "disgruntled" Differential Display user... 
I went through exactly what you are going through right now.  I've even
used the same "reverse Northern" technique because there were too many
"candidates" to screen.  And I hd the same problem you had with
everybody showing up on the dot blot.  We gave up on the project quickly
after that.  However, if you want to stick it out, I suggest a higher
stringency wash and hybridization.  I don't know if that will work, but
I think it is a good place to start.  In case you were wondering, we
went to another procedure called Representational Difference Analysis
(RDA) that served the same purpose as differential display (if you are
only comparing two populations).  The difference is RDA is cheaper to
run, less false positives, and no radioactive material until you are
ready to do the Northern blots, and no acrylamide gels...Let me know if
you are interested in the alternate methods, and I will find the
original article describing the procedure.  Also, I am curious to know
why you decided to do differential display (just to fulfill my own
curiosity).  Let me know if you have any other questions regarding
either method...God knows I spent enough time (almost too much) on
them...



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