First of all I'd like to thank all those who responded to my question.
My mailbox was swamped. I wish that I could respond to everyone personally
but that was not possible.
EVERYONE who responded suggested adding detergent (Tween, Triton, or
SDS) to the Clorox solution. This was something that we were not doing and
could solve much of the problem. Four respondents stated that they use full-
strength (100%) Clorox for 8-10 min with no adverse effects on the seed
germination. Two other respondents stated that the "freshness" of the bleach
could be a factor particularly in the summer and if the bottles have been in
storage a long time.
One respondent suggested that completely drying the seed before storage
(i.e. spread to dry under a laminar flow hood) significantly reduced their
fungal problems. And two respondants found that covering shaker flasks with
Parafilm reduced late fungal contamination in their cultures.
Antonio Franco (bf1rodri at cc.uco.es) is the person who first reported
the microwave sterilization technique. It is a bit unorthodoxed but I will
certainly try it and if it works will be a big time saver. I have asked him
to post the technique here on the Arabidopsis Board so that others can try it
as well. Basically small amounts of seed (50 seeds) are microwaved in sterile
plastic tubes. This would allow the convenience of having tubes of sterile
Again, I thank everyone who took the time to respond. I look forward
to meeting many of you in Columbus in August!
Melissa Melan mmelan at lucy.wellesley.edu
Department of Biological Sciences
Wellesley, MA 02181