rna fingerprinting help

schuchert at urz.unibas.ch schuchert at urz.unibas.ch
Fri Sep 24 03:45:26 EST 1993


dear netters
We are currently using the method of RNA fingerprinting to
display expression differences in the life stages of a marine
hydrozoan. In our hand the method described by Liang and Pardee
(1992, Science 257:967-971) did not work. It always produced a 
random pattern. However, the method of Welsh et al. (1992, NAR
20:4965-4970) did give excellent results.
But also the latter method leaves us with some problems. Out of 
the 15 tested primers only 7 worked (all approx. 60 % GC, 20 mer).
CAn anyone give an explanation for that or has anyone made the same
experience?
A further problem is the reproducibility of the cDNA step. If we 
have a good cDNA batch, we can take an aliquot and very well reproduce
the banding pattern by the PCR reaction. But in some cDNA batches
made with the same oligo, RNA etc, additional nonspecific bands
aas this problem been experienced by others too?

thanks heaps for all comments
Peter

Peter Schuchertnibas.ch
                                         



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