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Medium No.	1287
Medium	Modified D58 medium
Composition
Solution A* 440 ml   Solution B** 200 ml   Solution C*** 300 ml   Solution D**** 50 ml   Solution E***** 10 ml   pH 7.0-7.2       Mix solutions A, B and C, adjust pH to 7.0-7.2. Dispense the medium into suitable culture vessels under a stream of the N2/CO2 (80/20) mixed gas, seal with butyl rubber stoppers, and autoclave. After autoclaving, aseptically and anaerobically add filter-sterile solutions D and E to the medium using sterile syringes. The sterile syringe, filter and needle are flushed with nitrogen to remove the air before using. During preparation of the medium, precipitation may occur. But it not has much influence on the growth of the bacteria. The color of resazurin becomes colorless within several days after making the medium.                   *Solution A       (NH4)2SO4 1.3 g       KH2PO4 1.43 g       K2HPO4·3H2O 7.2 g       FeSO4·7H2O 1.1 mg       Yeast extract 4.5 g       Resazurin 1 mg       Distilled water 440 ml                   **Solution B       MgCl2·6H2O 2.6 g       CaCl2·2H2O 0.13 g       Distilled water 200 ml     Completely dissolve ingredients in distilled water.                   ***Solution C       Sodium glycerophosphate 6 g       Distilled water 300 ml                   ****Solution D       Cellobiose 5 g       Distilled water 50 ml     Completely dissolve ingredient in hot distilled water, and remove air by bubbling with nitrogen gas.                   *****Solution E       Glutathione 0.25 g       Distilled water 10 ml     Completely dissolve ingredient in distilled water, and remove air by bubbling with nitrogen gas.
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