Bacillus subtilis subsp. subtilis (Ehrenberg) Cohn
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. From a tube of #415 broth (5 to 6 ml), withdraw approximately 0.5 with a sterile pipette; add this medium to the vial to rehydrate the entire pellet.
3. Aseptically transfer the contents of the vial into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a #415 agar slant and/or plate.
5. Incubate all tubes and plate at 26°C for 48 hours.
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