Re-using hybridisation solutions containing double-stranded

Roger Palfree MCRP000 at MUSICA.MCGILL.CA
Tue Feb 28 18:41:20 EST 1995


In article <01HNG6291XDUA5U66B at MPGARS.DESY.DE>                          tes:
DREWES at MPASMB.DESY.DE writes:
>Has anybody ever tried re-using plaque hybridisation solutions containing
>32P-labeled double stranded probes? It would mean a significant reduce in
>radioactive waste and expensive stuff like salmon sperm DNA and deionized
>formamide. I am thinking about simply storing the used solution in the freezer
>and then denature the DNA and the probe again by boiling for ten minutes before
>the next use.

If you are hybridizing in solutions containing formamide, there should
be no difficulty denaturing the probe again by heating.  I have done
this and reused probe successfully.  If you apply the formula for Tm
of your probe in other hybridization buffers, I think you will find
that, in theory, boiling should work in many cases.  Just bear in mind
if you do this, that a negative result will leave you wondering if
your reused probe was any good after all!

Roger Palfree




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