verifying DDPCR positives

Steven Sullivan sullivan at gwis2.circ.gwu.edu
Sat May 17 00:04:52 EST 1997



I have a handful (5) of cloned fragments that appear to be differentially
displayed in my target tissues, but only a limited amount of RNA and
tissue (and time !).  Northerns are out of the question (not enough RNA); 
what other methods might I use to quickly verify the differential display? 
I tried RT-PCR using internal primers, which indicated my fragments are
false positives;  but it occurs to me that RT-PCR (which was not
quantitative in this case)  might be *too* sensitive, obscuring all but
the most radical differences in expression.  What to try next? Dot blots?
RNase protection? Some sort of quantitative RT-PCR? 





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