Michael J. Prigge
prigge at darkwing.uoregon.edu
Tue Dec 17 17:22:19 EST 1996
In article <596s7p$8ps10 at ncisun1-nf0.ncifcrf.gov>, pnh at ncifcrf.gov (Paul N
> Bernard Murray (bernard at elsie.nci.nih.gov) wrote:
> > From what I understand moisture is essential for the sterilisation
> > process. Another poster described a microwave based sterilisation
> > unit that involved the generation of steam. I don't think that
> > simply microwaving dry glassware in the oven has any effect.
> I don't think so either.
I beg to differ on this point... a method I have used to sterilize
Arabidopsis seed is essentially dry. I nuked 15 ml polyprop tube
with a few hundred seed twice for five min each time. At the end
of the dose, the tube was kinda soft but the seed was ~100% viable
AND NO FUNGUS (or anything else) GREW when plated.
Now for the potential caveats: I plated the seed onto kanamycin
or hygromycin plates; and there was a beaker of water in the
microwave--to soak up unquenched microwaves(?)--that was replaced as
it began to boil. Without sterilization, though, kan and hyg plates
are overrun with fungii before germination.
Evidently, the water content of the bacteria and spores and lack
thereof in the dehydrated seed embryos contributes to the
differential killing. I know that, if weed contamination is a
problem for you, longer runs take care of the Arabidopsis too. :)
Michael Prigge prigge at darkwing.uoregon.edu
Institute of Molecular Biology University of Oregon
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