PCR products in Southerns

Shiao Y. Wang sywang at whale.st.usm.edu
Wed Jul 27 14:28:39 EST 1994


Judy Broom (judy at sanger.otago.ac.nz) wrote:

: I would like to use PCR products as probes in Southerns (using 
: the Boehringer Random Primed DNA Labeling Kit).  What purification 
: of the PCR product is necessary?  Someone recommended the Bio Rad Pr
: ep-A-Gel system to me - would a crude ethanol precipitation do in
: stead?  (I tried probing with NO purification, and it sort of worked... )

Why don't you just make your probe by PCR directly? Gibco/BRL sells a kit
for doing so but the components are those used for normal PCR. That way you
can skip the purification step altogether. I'm trying this but am having
difficulty getting good incorporation. Will post a success story soon I hope.

If you want to gel-purify the DNA fragment, BioRad's Prep-A-Gene kit works
very nicely (I've had better luck with it than GeneClean). The DNA size
limit still holds though (400 bp I think). Good luck.

Shiao Wang
University of Southern Mississippi




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