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Medium Detail Information

Medium No.
927 
Medium
Medium for iron-using methanogen 
Composition
 HEPES23.8g
 NaCl19g
 MgCl2·6H2O2.6g
 CaCl2·2H2O0.15g
 Na2SO44g
 NH4Cl0.25g
 KH2PO40.2g
 KCl0.5g
 Trace elements solution*1ml
 Vitamin solution**1ml
 NaHCO32.52g
 Iron granule (1-2 mm)***150g
 Cysteine-HCl1g
 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
   FeCl3·6H2O1.35g
   MnCl2·4H2O0.1g
   CoCl2·6H2O0.024g
   CaCl2·2H2O0.1g
   ZnCl20.1g
   CuCl2·2H2O0.025g
   H3BO30.01g
   Na2MoO4·2H2O0.024g
   NaCl1g
   NiCl2·6H2O0.12g
   Na2SeO40.004g
   Na2WO40.004g
   KAl(SO4)2·12H2O0.02g
   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
   Biotin2mg
   Folic acid2mg
   Pyridoxine-HCl10mg
   Thiamine-HCl5mg
   Riboflavin5mg
   Nicotinic acid5mg
   Ca-pantothenate5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L
       
  ***Alfa Aesar A Johnson Matthey Company, Massachusetts, USA.
 
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