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Medium Detail Information

Medium No.	1298
Medium	Hydrogen-using marine methanogen medium
Composition
KP buffer* 5 ml   MgCl2·6H2O 0.75 g   CaCl2·2H2O 0.15 g   NH4Cl 0.3 g   NaNO3 0.3 g   NaCl 30 g   Sodium acetate 0.16 g   Bacto Yeast Extract (Difco) 0.2 g   Trace elements solution** 2 ml   Vitamin solution*** 2 ml   Resazurin 1 mg   Na2CO3 1 g   Cysteine-HCl 0.35 g   Na2S·9H2O 0.35 g   Distilled water 1 L   pH unadjusted       Mix ingredients except KP buffer, vitamin solution, Na2CO3, cysteine-HCl and Na2S·9H2O. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave KP buffer, cysteine-HCl (5% solution) and Na2S·9H2O (5% solution) under a N2 atmosphere. Sterilize vitamin and 10% Na2CO3 solutions by filtration. Prior to inoculation, aseptically and anaerobically add KP buffer, vitamin, Na2CO3, cysteine-HCl and Na2S·9H2O solutions. Pressurize the inoculated vessels to 150 kPa with H2/CO2 (80/20).                   *KP buffer       KH2PO4 119 g       K2HPO4 21 g       Distilled water 1 L     Autoclaved under N2.                   **Trace elements solution       Nitrilotriacetic acid (NTA) 12.8 g       FeCl3·6H2O 1.35 g       MnCl2·4H2O 0.1 g       CoCl2·6H2O 0.024 g       CaCl2·2H2O 0.1 g       ZnCl2 0.1 g       CuCl2·2H2O 0.025 g       H3BO3 0.01 g       Na2MoO4·2H2O 0.024 g       NaCl 1 g       NiCl2·6H2O 0.12 g       Na2SeO4 0.004 g       Na2WO4 0.004 g       KAl(SO4)2·12H2O 0.02 g       Distilled water 1 L     First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.                   ***Vitamin solution       Biotin 2 mg       Folic acid 2 mg       Pyridoxine-HCl 10 mg       Thiamine-HCl 5 mg       Riboflavin 5 mg       Nicotinic acid 5 mg       Ca-pantothenate 5 mg       p-Aminobenzoic acid 1 mg       Vitamin B12 0.01 mg       Distilled water 1 L
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