S1 nuclease + PCR to verify spliced mRNA?

Stefi~ xxxgifalone at tin.it
Mon Jul 2 13:17:22 EST 2001


<vuvu at freemail.it> wrote in message 3B40A968.9EC8DF30 at freemail.it...
> I wonder if to verify the existence of a deletion in mRNA one could
> hybridize it with a DNA probe, followed by S1 nuclease, then isolate the
>
> ssDNA probe, go to dsDNA and do a PCR  , therefore skipping use of
> radioactivity

how could u have your nuclease S1-digested hybrid DNA-mRNA converted in
dsDNA. You'll have several fragments (introns + 1) not ligated. And if you
want to amplify yaour fragments you'll have to design and use two primers
for each exon you want to be amplified. If I'm wrong, tell me so I'll
increase my knowledge in PCR technique.
Stefano
gifalone at tin.it

http://web.tiscalinet.it/gifalone
ICQ 110419911





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