Microbiology List: FW: Bacillus sporulation

Yersinia 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 
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.

Yersinia<----oh, it's been TOO long!

Mycelium is yourcelium.


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