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Medium Detail Information

Medium No.
911 
Medium
Marine H2-using Methanogen Medium 
Composition
 KP7 buffer*5ml
 MgCl2·6H2O3.05g
 CaCl2·2H2O0.15g
 NH4Cl0.54g
 Na2SO40.3g
 NaCl30g
 Bacto Yeast Extract (Difco)0.02g
 Sodium acetate0.8g
 Trace elements solution**2ml
 Vitamin solution***2ml
 Resazurin1mg
 Na2CO31.5g
 Na2S·9H2O0.5g
 Cysteine-HCl0.5g
 Distilled water1L
 pH unadjusted 
  Mix ingredients except KP7 buffer, vitamin solution, Na2CO3, Na2S·9H2O and cysteine-HCl. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave KP7 buffer, Na2S·9H2O (5% solution) and cysteine-HCl (5% solution) under a N2 atmosphere. Sterilize vitamin and 10% Na2CO3 solutions by filtration. Prior to inoculation, aseptically and anaerobically add KP7 buffer, vitamin, Na2CO3 and Na2S·9H2O solutions. Pressurize the inoculated vessels to 150 kPa with H2/CO2.
       
  *KP7 buffer
   KH2PO453g
   K2HPO4106g
   Distilled water1L
  Autoclaved under N2.
       
  **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 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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