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Medium Detail Information

Medium No.
880
Medium
Methanoculleus Medium
Composition
 NH4Cl0.25g
 KCl0.34g
 MgCl2·6H2O4g
 MgSO4·7H2O3.5g
 CaCl2·2H2O0.14g
 K2HPO40.14g
 NaCl6g
 Trace elements solution*10ml
 Vitamin solution**10ml
 Bacto Yeast Extract (Difco)2g
 Polypeptone2g
 Sodium acetate1g
 Resazurin1mg
 NaHCO35g
 Cysteine-HCl0.5g
 Na2S·9H2O0.5g
 Distilled water1L
  Mix ingredients except for vitamin solution, NaHCO3, cysteine-HCl and Na2S·9H2O, and autoclave under H2/CO2 (80/20) condition. Separately autoclave cysteine-HCl and Na2S·9H2O as a 5% solution under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin and NaHCO3 solutions, and cysteine-HCl and Na2S·9H2O solutions, and pressurize inoculated bottles to 150 kPa with H2/CO2.
       
  *Trace elements solution
   Nitrilotriacetic acid (NTA)4.1g
   FeCl3·6H2O0.36g
   MnCl2·4H2O0.09g
   CoCl2·6H2O0.15g
   CaCl2·2H2O0.018g
   ZnCl20.09g
   H3BO30.017g
   Na2MoO4·2H2O0.009g
   NiCl2·6H2O0.018g
   Distilled water1L
  First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add 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
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