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Medium No.	837
Medium	Methanosarcina Medium
Composition
KH2PO4 0.75 g   K2HPO4 0.75 g   MgCl2·6H2O 0.36 g   NH4Cl 1 g   Bacto Yeast Extract (Difco) 0.1 g   Hipolypepton* 0.1 g   Sodium acetate 2.5 g   Trimethylamine-HCl 8 g   Trace mineral solution** 9 ml   Trace vitamins*** 10 ml   NaHCO3 0.8 g   Na2S·9H2O 0.5 g   Cysteine-HCl 0.5 g   Resazurin 1 mg   Distilled water 981 ml     Mix ingredients except trace vitamins, Na2S·9H2O and cysteine-HCl, and autoclave under a N2/CO2 (80/20) atmosphere. Separately autoclave Na2S·9H2O and cysteine-HCl as 5% solutions under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin solution to the medium. Prior to inoculation, add the sterile Na2S·9H2O and cysteine-HCl solutions, and adjust pH to 6.8.                   *Wako Pure Chemicals Ind. Ltd., Osaka, Japan.                   **Trace mineral solution       Nitrilotriacetic acid 4.5 g       FeCl2·4H2O 0.4 g       MnCl2·4H2O 0.1 g       CoCl2·6H2O 0.17 g       ZnCl2 0.1 g       CaCl2·2H2O 0.02 g       H3BO3 0.019 g       Na2MoO4·2H2O 0.01 g       NiCl2·6H2O 0.03 g       Distilled water 1 L   Adjust pH to 7.0 with 1 N KOH                     ***Trace vitamins       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       Vitamin B12 0.1 mg       p-Aminobenzoic acid 5 mg       Lipoic acid 5 mg       Distilled water 1 L
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