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Medium Detail Information

Medium No.
1083
Medium
Methanobacterium alcaliphilum medium
Composition
 NH4Cl1g
 MgCl2·6H2O0.1g
 CaCl2·2H2O0.1g
 K2HPO40.4g
 Bacto Yeast Extract (Difco)2g
 Polypeptone2g
 Resazurin1mg
 Carbonate mixture*210ml
 Tris-HCl buffer**10ml
 Vitamin solution***10ml
 Trace elements solution****10ml
 Cysteine-HCl0.3g
 Na2S·9H2O0.3g
 Distilled water760ml
  Mix ingredients except for carbonate mixture, Tris-HCl buffer, vitamin solution, cysteine-HCl and Na2S·9H2O, and autoclave under a H2 atmosphere. Carbonate mixture, Tris-HCl buffer, cysteine-HCl (3% solution) and Na2S·9H2O (3% solution) are separately autoclaved under a N2 atmosphere. Prior to inoculation, add filter-sterile vitamin solution, carbon mixture, Tris-HCl buffer, cysteine-HCl and Na2S·9H2O. Pressurize the inoculated vessels to 150 kPa with H2.
       
  *Carbonate mixture
   NaHCO310g
   Na2CO30.5g
   Distilled water210ml
  Mix ingredients and autoclave under a N2 atmosphere.
       
  **Tris-HCl buffer
  Dissolve 2 M Tris, adjust pH to 8.4 with HCl, and autoclave under a N2 atmosphere.
       
  ***Vitamin solution
   Biotin2mg
   Folic acid2mg
   Pyridoxine-HCl10mg
   Thiamine-HCl5mg
   Riboflavin5mg
   Nicotinic acid5mg
   Ca-pantothenate5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L
       
  ****Trace elements solution
   2Na-EDTA0.64g
   MgSO4·7H2O6.2g
   MnSO4·4H2O0.55g
   NaCl1g
   FeSO4·7H2O0.1g
   CoCl2·6H2O0.17g
   CaCl2·2H2O0.13g
   ZnSO4·7H2O0.18g
   CuSO40.05g
   KAl(SO4)2·12H2O0.018g
   H3BO30.01g
   Na2MoO4·2H2O0.011g
   NiCl2·6H2O0.025g
   Na2SeO40.004g
   Na2WO40.004g
   Distilled water1L
  Dissolve the 2Na-EDTA in 500 ml of distilled water, and after adding the salts, make up to 1 L.
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