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Medium Detail Information

Medium No.
837
Medium
Methanosarcina Medium
Composition
 KH2PO40.75g
 K2HPO40.75g
 MgCl2·6H2O0.36g
 NH4Cl1g
 Bacto Yeast Extract (Difco)0.1g
 Hipolypepton*0.1g
 Sodium acetate2.5g
 Trimethylamine-HCl8g
 Trace mineral solution**9ml
 Trace vitamins***10ml
 NaHCO30.8g
 Na2S·9H2O0.5g
 Cysteine-HCl0.5g
 Resazurin1mg
 Distilled water981ml
  Mix ingredients except trace vitamins, Na2S·9H2O and cysteine-HCl, and autoclave under a N2/CO2 (80/20) atmosphere. Separately autoclave Na2S·9H2O and cysteine-HCl as 5% solutions under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin solution to the medium. Prior to inoculation, add the sterile Na2S·9H2O and cysteine-HCl solutions, and adjust pH to 6.8.
       
  *Wako Pure Chemicals Ind. Ltd., Osaka, Japan.
       
  **Trace mineral solution
   Nitrilotriacetic acid4.5g
   FeCl2·4H2O0.4g
   MnCl2·4H2O0.1g
   CoCl2·6H2O0.17g
   ZnCl20.1g
   CaCl2·2H2O0.02g
   H3BO30.019g
   Na2MoO4·2H2O0.01g
   NiCl2·6H2O0.03g
   Distilled water1L
 Adjust pH to 7.0 with 1 N KOH 
       
  ***Trace vitamins
   Biotin2mg
   Folic acid2mg
   Pyridoxine-HCl10mg
   Thiamine-HCl5mg
   Riboflavin5mg
   Nicotinic acid5mg
   Ca-pantothenate5mg
   Vitamin B120.1mg
   p-Aminobenzoic acid5mg
   Lipoic acid5mg
   Distilled water1L
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