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Medium Detail Information

Medium No.	860
Medium	Methanosarcina acetivorans Medium
Composition
NaCl 23.4 g   MgSO4·7H2O 9.45 g   Na2CO3 5 g   NH4Cl 1 g   KCl 0.8 g   CaCl2·2H2O 0.14 g   Na2HPO4 0.6 g   Bacto Yeast Extract (Difco) 1 g   Trace elements solution* 10 ml   Vitamin solution** 10 ml   Methanol 6.5 ml   Resazurin 1 mg   Cysteine-HCl 0.3 g   Na2S·9H2O 0.3 g   Distilled water 1 L     Mix ingredients except methanol, vitamin solution, cysteine-HCl, and Na2S·9H2O. Flush with N2/CO2, and adjust pH to 7.0 with HCl. Dispense the medium into suitable culture vessels under a stream of N2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave cysteine-HCl and Na2S·9H2O (5% solution) under a N2 atmosphere. Sterilize methanol and vitamin solution by filtration. Prior to inoculation, aseptically and anaerobically add vitamin, cysteine-HCl and Na2S·9H2O solutions.                   *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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