verifying DDPCR positives
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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