Thermus aquaticus Brock and Freeze
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2. Using a single tube of #461 broth (5 to 6 ml), withdraw approximately 0.6 ml with a Pasteur or 1.0 ml pipette. Rehydrate the entire pellet.
3. Aseptically transfer 0.3 ml of this suspension back into the broth tube. Mix well.
4. Use the remaining 0.3 ml of the suspension to inoculate a slant of #461 agar.
5. Incubate tubes at 70oC. For best results, incubate tubes within a closed jar with a moistened paper towel added to maintain humidity and prevent desiccation.
6. Once growth is obtained in broth, plates can be inoculated with approximately 0.2 ml per plate. Better growth is achieved on plates when incubated at 60oC rather than 70oC. Plates should be incubated in a jar as described above or taped shut to prevent desiccation.
Yellow growth should be visible in broth and slant tubes after 24 to 48 hours of incubation. At 1000X magnification, cells are long, regular rods occurring singly or occasionally as end-to-end pairs.
After 48 hours, the colonies on plates are circular with a convex elevation, entire margin, and a smooth surface. They are dark yellow in color and can be up to 1 mm in diameter.
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Brock TD, Freeze H. Thermus aquaticus gen. n. and sp. n., a nonsporulating extreme thermophile. J. Bacteriol. 98: 289-297, 1969. PubMed: 5781580
Skerman VB, et al. Approved lists of bacterial names. Int J Syst Bacteriol 30: 225-420, 1980.
Roberts RJ. Restriction and modification enzymes and their recognition sequences. Nucleic Acids Res. 13: r165-r200, 1985. PubMed: 2987885
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