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Medium Detail Information

Medium No.	406
Medium	Methanobacterium Medium
Composition
KH2PO4 0.3 g   K2HPO4 0.3 g   (NH4)2SO4 0.3 g   NaCl 0.6 g   MgSO2·7H2O 0.13 g   FeSO2·7H2O 2 mg   CaCl2·2H2O 8 mg   Bacto Yeast Extract (Difco) 2 g   Trypticase Peptone (BBL) 2 g   Sodium acetate 2.5 g   Sodium formate 2.5 g   Trace minerals* 10 ml   Trace vitamins** 10 ml   NaHCO3 5 g   Na2S·9H2O 0.5 g   Cysteine-HCl 0.5 g   Resazurin 1 mg   Distilled water 1 L     Mix ingredients except NaHCO3, cysteine-HCl and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of H2/CO2 (80/20), and add NaHCO3 to the medium. Dispense the medium into suitable culture vessels (e.g. 20 ml of the medium in 120 ml serum bottles) under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave cysteine-HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine-HCl and Na2S·9H2O solutions. Pressurize the inoculated vessels to 200 kPa with H2/CO2 (80/20).                   *Trace minerals       Nitrilotriacetic acid 1.5 g       MgSO4·7H2O 3 g       MnSO4·xH2O 0.5 g       NaCl 1 g       FeSO4·7H2O 0.1 g       CoSO4·7H20 0.1 g       CaCl2·2H2O 0.1 g       ZnSO4·7H2O 0.1 g       CuSO4·5H2O 0.01 g       AlK(SO4)2 0.01 g       H3BO3 0.01 g       Na2MoO4·2H2O 0.01 g       Distilled water 1 L     Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.                   **Trace vitamins       Biotin 2 mg       Folic acid 2 mg       Pyridoxine-HCl 10 mg       Thiamine-HCl 5 mg       Riboflavin 5 mg       Nicotinic acid 5 mg       Calcium 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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