S1 nuclease + PCR to verify spliced mRNA?

vuvu at freemail.it vuvu at freemail.it
Mon Jul 2 12:04:00 EST 2001


Hi
I found some deletions in a mRNA by rt-pcr; now I'm reading that there
are cases in which loops in RNA can be jumped (see template switching in
med line) and I'd like to point out to the group the importance of the
new reverse transcriptases working at higher temperatures

Anyway I'm in the situation that I need to verify if the deletions I'm
observing are real

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

thanks
Vuvu

---

---




More information about the Methods mailing list