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Medium Detail Information

Medium No.
Medium for iron-using methanogen
 Trace elements solution*1ml
 Vitamin solution**1ml
 Iron granule (1-2 mm)***150g
 Distilled water1L
  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)12.8g
   Distilled water1L
  First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add other minerals. Final pH 7.0.
  **Vitamin solution
   Folic acid2mg
   Nicotinic acid5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L
  ***Alfa Aesar A Johnson Matthey Company, Massachusetts, USA.