PCR probes

Brian Foley brianf at med.uvm.edu
Thu Apr 14 18:18:24 EST 1994


Shiao Y. Wang (sywang at whale.st.usm.edu) wrote:
: : > 
: : > 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).

: 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

	No price information, but another significant feature of a PCR
reaction for labeling probe is that you can specifically label any
piece of a plasmid, without labelling everything (or gel-purifying
a small band for random primers).
	For example this is good if your plasmid would cross-hybridize
to things.  Or if you are looking for alernative splicing or some
other small change.

--
********************************************************************
*  Brian Foley               *     If we knew what we were doing   *
*  Molecular Genetics Dept.  *     it wouldn't be called research  *
*  University of Vermont     *                                     *
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