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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 single tube of #435 broth (5 to 6 ml), withdraw approximately 0.5 to 1.0 ml with a Pasteur or 1.0 ml pipette and use to rehydrate the entire pellet.
3. Aseptically transfer the rehydrated pellet back into the broth tube. Mix well.
4. Use several drops of this suspension to inoculate an additional broth tube, agar slant and/or a plate.
5. Incubate the tubes and plate at 30°C for 48 to 72 hours. If growth is not heavy on agar medium, make additional transfers from the incubated broth at this time.
Additional information on this culture is available on the ATCC web site at www.atcc.org.
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Nakayama O, Yanoshi M. Spore-bearing lactic acid bacteria isolated from rhizosphere. I. Taxonomic studies on Bacillus laevolacticus nov. sp. and Bacillus racemilacticus nov. sp.. J. Gen. Appl. Microbiol. 13: 139-153, 1967.
Hatayama K, et al. Tuberibacillus calidus gen. nov., sp. nov., isolated from a compost pile and reclassification of Bacillus naganoensis Tomimura et al. 1990 as Pullulanibacillus naganoensis gen. nov., comb. nov. and Bacillus laevolacticus Andersch et al. 1994 as Sporolactobacillus laevolacticus comb. nov. Int. J. Syst. Evol Microbiol. 56(Pt 11): 2545-2551, 2006. PubMed: 17082388