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Medium Detail Information

Medium No.	927
Medium	Medium for iron-using methanogen
Composition
HEPES 23.8 g   NaCl 19 g   MgCl2·6H2O 2.6 g   CaCl2·2H2O 0.15 g   Na2SO4 4 g   NH4Cl 0.25 g   KH2PO4 0.2 g   KCl 0.5 g   Trace elements solution* 1 ml   Vitamin solution** 1 ml   NaHCO3 2.52 g   Iron granule (1-2 mm)*** 150 g   Cysteine-HCl 1 g   Distilled water 1 L     Mix ingredients, except vitamin solution, NaHCO3, iron granule and cysteine-HCl. Dispense the medium into suitable culture vessels under a stream of N2/CO2 (80/20), seal with butyl rubber stoppers, and autoclave for 20 min at 121°C. Sterilize vitamin and 8.4% NaHCO3 solutions by filtration. 5% Cysteine-HCl solution and iron granule are separately sterilized by autoclaving for 20 min at 121°C under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add vitamin, NaHCO3, iron granule and cysteine-HCl.                   *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 other 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                   ***Alfa Aesar A Johnson Matthey Company, Massachusetts, USA.
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