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
transcription step?

, 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..

Martin
> If so, please me kno as soon as you can.
> Thanks.

-- 
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