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
for you:
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.
Infectionately,
Yersinia<----oh, it's been TOO long!
Mycelium is yourcelium.
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