Microbiology List: FW: Bacillus sporulation
yersinia at cybernex.net
Tue Aug 8 17:19:12 EST 2000
Walter Blank writes,
<I'm trying to find procedures for inducing sporulation in Bacillus (B.t.
or subtilis), separating out vegetative cells, and enumerating the
spores by cfu's (so I guess they'd have to be effectively germinated).
If anyone could pass along or point me in the direction of favorite
methods I'd appreciate it.>
If my memory serves me correctly (it's been awhile!), this ought to do it
Grow your B.subtilis (or thuringiensis) out on AK Sporulating Agar.
Harvest the growth into test tubes and heat shock them at 80C for 15
minutes. Centrifuge (sorry, I don't recall for how long or how many
rpm's; maybe someone else can help here). Pour off the supernatant; the
spores are in the pellet. Add alcohol (ethanol) I think, as suspension
liquid. When the spores are in suspension, you can make dilutions and
plate out for enumeration.
Yersinia<----oh, it's been TOO long!
Mycelium is yourcelium.
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