RT-PCR using S1 Nuclease
Martin D. Leach
leach at bu.edu
Sat Mar 18 12:18:27 EST 1995
In article <3kd2lm$olo at steele.ohsu.EDU>, "Petra M. Jakobs, Ph.D."
<jakobsp at ohsu.edu> wrote:
> From: Philip S. Ruban Sahaayar at ohsu.edu
> I am trying to do RT-PCR, using S1 nuclease.
How about using a reverse transcriptase? AMV or MuLV.
Basically, I first extract
> my RNA and throw the probe of my interest into the total RNA. Then cut
> with S1 nuclease and
> amplyfy the hybrid.
How? Using S1 will digest all the single stranded DNA/RNA...Where are the
primers sticking to? What is the primer you are using for the reverse
, using PCR.
> Could anyone know the protocol for this method?
There is a S1-nuclease digestion protocol found in Maniatis (the bible)
but this does not involve any PCR...You just run the result of the
digestion on an acrylamide gel..
> If so, please me kno as soon as you can.
..... Martin Leach Email:leach at bu.edu
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