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Medium Detail Information

Medium No.
249
Medium
Composition
 Na2HPO4·12H2O4.2g
 KH2PO41.5g
 Na2CO34g
 MgCl2·6H2O0.4g
 NH4Cl0.6g
 Yeast extract1g
 Bacto Tryptone (Difco)1g
 Na2WO4·2H2O0.4mg
 Wolfe's modified mineral solution*50ml
 VFA**1ml
 Distilled water738ml
 pH 6.5 
  Adjust pH to 6.5 with 10 N KOH and add the following medium.
   0.1% Resazurin solution1ml
   myo-Inositol1mg
   Vitamin mixture***10ml
   Cellobiose0.2g
   Cellulose****200ml
  Heat the above mixed solution at 60℃ for 5 to 10 min and then bubble with N2 gas. And then add the following ingredients.
   Cysteine0.5g
   Na2S·9H2O0.5g
   Agar (if needed)15g
  Under anaerobic condition, the medium is autoclaved at 100℃ for 10 min.
       
  *Wolfe's modified mineral solution
   Nitrilotriacetic acid (NTA)1.5g
   MgSO4·7H2O3g
   NaCl1g
   MnSO4·H2O0.5g
   FeSO4·7H2O0.1g
   Co(NO3)2·6H2O0.1g
   CaCl20.1g
   ZnSO4·7H2O0.1g
   CuSO4·5H2O0.01g
   Boric acid0.01g
   Na2MoO4·2H2O0.01g
   Na2SeO30.001g
  Nitrilotriacetic acid is suspended in 500 ml water, and dissolved by titrating with 2 N KOH until its pH stabilize at 6.5. The rest of the ingredients are added and dissolved. Finally total volume is adjusted to 1 L.
       
  **VFA
   Acetic acid17ml
   Propionic acid6ml
   Butyric acid4ml
   iso-Butyric acid1ml
   n-Valeric acid1ml
   iso-Valeric acid1ml
   DL-α-Methylbutyric acid1ml
       
  ***Vitamin mixture
   Biotin0.2mg
   Ca-pantothenate40mg
   Folic acid0.2mg
   myo-Inositol200mg
   Niacin40mg
   p-Aminobenzoic acid20mg
   Pyridoxine-HCl40mg
   Riboflavin40mg
   Thiamin-HCl20mg
   Choline chloride100mg
   Cyanocobalamin2μg
   Distilled water1L
       
  ****3% Cellulose suspension which is ball-milled for 3 days.
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