Method for sterilizing small seeds

Andreas Wistuba (andreas.wistuba@rhein-neckar.de)
Fri, 08 Mar 1996 06:37:02 GMT

After I tried a lot of techniques described in various papers and
reviews to sterilize very small seeds such as Orchid seeds or seeds of
Drosera, Pinguicula Genlisea, Utricularia....., I found a nice and
easy method which at least so far I did not find in any publication.

Just sterilize the seeds in bleach (as usual) in an Eppendorf tube.
After sterilizing just remove the liquid by pressing the opening of a
syringe needle strongly against the tube wall creating a very narrow
filter. You can then remove the liquid with the syringe and add
sterile water with an Eppendorf pipette. The whole is repeated two
times. The advantage compared with other techniques involving
centrifugation is that it does not matter whether the seeds swim or
sink (very light seeds immediately move to the surface again when the
centrifuge stops....). Compared to the usuall filtration methods such
as filtrating via a Buchner funnel on a filter paper the advantage is
that if done carefully you do not lose any seeds. I sterilized very
small amounts without any trouble. The sterilization is far better
than in case of methods involving little envelopes of filter paper to
hold the seeds during bleaching.

After sterilization the seeds are sown by removing them in the water
with an Eppendorf pipette and sprayed on the agar surface. If
neccesary fresh water can be added a few times to enable removing all
the seeds from the tube.

Bye,

Andreas
Andreas Wistuba; Mudauer Ring 227; 68259 Mannheim; Germany
Phone: +49-621-705471 / +49-621-7152027
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