PCR probes

Molapo Qhobela 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
>: wrote:

>: > 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).

Hmmm,

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.

Regards,

Molapo



>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
>real numbers?

>Shiao Wang
>University of Southern Mississippi


Molapo Qhobela PhD                 |   Department of Microbiology
Internet : molapo at micro.uct.ac.za  |   University of Cape Town
Tel: +27 (21) 650-3263             |   Private Bag, Rondebosch 7700
Fax: +27 (21) 650-4023             |   South Africa




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