Caldicellulosiruptor acetigenus (Nielsen et al.) Onyenwoke et al.
BAA-1149 ™
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2. If needed exchange the gas in the test tube for 80% N2-20% CO2.
3. Open vial according to enclosed instructions.
4. Under anaerobic conditions, withdraw 0.5 ml of recommended broth from a single test tube (5 to 6 ml) and rehydrate the entire vial contents.
5. Aseptically, using a 1.0 ml syringe tipped with 22-gauge needle, withdraw the cell suspension from the vial and transfer it to the broth. Plate 0.1 ml of the inoculated culture onto a non-selective medium and incubate aerobically at 30oC. Use 0.1 ml of the inoculated culture to inoculate a non-selective aerobic broth. Incubate the inoculated tubes at 30oC.
6. Growth should be detected in the #2587 broth in 24. There should be no growth detected on the aerobic plate or broth.
ANAEROBIC CONDITIONS:
a. Resazurin is a commonly used redox indicator that is pink when the redox potential is above 50 mv., and colorless when the redox potential is below 110 mv. i.e. highly reducing. Most strict anaerobes require this low redox potential for optimum growth.
b. To obtain a fully reduced medium, it is necessary that the medium be anoxic and that a reducing agent be added. Common reducing agents are sodium sulfide, cysteine, dithiothreitol, and titanium citrate.
c. Syringes can be made anaerobic by one of two methods. 1. Displace the dead space in the syringe with a sterile
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Onyenwoke RU, et al. Reclassification of Thermoanaerobium acetigenum as Caldicellulosiruptor acetigenus comb. nov. and emendation of the genus description. Int. J. Syst. Evol. Microbiol. 56(Pt 6): 1391-1395, 2006. PubMed: 16738119