RT-PCR using S1 Nuclease

[Shahram Mori]

Martin D. Leach (leach at bu.edu) wrote:
: In article <3kd2lm$olo at steele.ohsu.EDU>, "Petra M. Jakobs, Ph.D."
: <jakobsp at ohsu.edu> wrote:

:  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?
: -- 
: .....          Martin Leach                Email:leach at bu.edu 
:    _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
:    / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
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Well, you can't do PCR on the S1 reaction anyway! You have to isolate the DNA
using spin columns or something. That will keep S1 away and allow the PCR 
to continue.
Cheers
--
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||
88003