Bacillus Subtillus culture

dwimmer at smartnet.net dwimmer at smartnet.net
Tue Nov 21 21:14:50 EST 1995

Derek & Nikki ALBERT wrote:
> Could anyone tell me the best way to culture and then store this
> bacteria in any numbers
> Regards Derek

1. Get your pure culture and inoculate a 100ml flask of an enriched 
broth media. Incubate 24hr at 37dC on a shaker so you get a good, 
turbid solution.
2. To eight flasks with 250mL each of a "sporulation broth" which I 
think has low carbohydrates, inoculate with 10mL of the culture.
3. Shake n bake at R.T. for a couple of weeks. There will be a film 
on the top. Gram stain should show 90%+ sporulation.
4. Spin down the stuff to get a nice thick tan paste.
5. Bring up to about 50mL with sterile saline.
6. Heat shock at about 60dC for a half hour or so. Store in the 
This will give a culture containing about 8 logs per 0.1mL and 
should last at least a couple of years.

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