molapo at micro.uct.ac.za
Fri Apr 15 10:25:53 EST 1994
In article <2ojrkq$jp0 at server.st.usm.edu> sywang at whale.st.usm.edu (Shiao Y. Wang) writes:
>From: sywang at whale.st.usm.edu (Shiao Y. Wang)
>Subject: Re: PCR probes
>Date: 14 Apr 1994 16:39:54 GMT
>Martin Leach (leach at mbcrr.harvard.edu) wrote:
>: In article <1994Apr13.125331.5635 at leeds.ac.uk>, futers at bpxtal.leeds.ac.uk
>: > Hi netters,
>: > Has anyone performed a PCR reaction in the presence of a 32P dNTP and
>: > then used the product to probe a Southern blot?
>: > If there is any interest I will post a summary.
>: > Many thanks,
>: > Simon (futers at biovax.leeds.ac.uk)
>: I believe many people do this - although i prefer the random priming method
>: with hexamers (only takes 30mins).
Why not use DIG, incorporating DIG dUTP in the PCR reaction is no big deal ...
and we routinely use PCR labeled products to probe a Southern blot.
>Interesting question. Both produce hot probes that work well but PCR may
>have some advantages. E.g., PCR should produce far more labeled material and
>would be less expensive to make. The cost aspect may not be true because
>I'm thinking of random priming kits (around $5/reaction) vs components for
>PCR (around $1/reaction), not including labeled nucleotide. Anybody have
>University of Southern Mississippi
Molapo Qhobela PhD | Department of Microbiology
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