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Medium Detail Information

Medium No.
 Yeast extract1g
 Bacto Tryptone (Difco)1g
 Wolfe's modified mineral solution*50ml
 Distilled water738ml
 pH 6.5 
  Adjust pH to 6.5 with 10 N KOH and add the following medium.
   0.1% Resazurin solution1ml
   Vitamin mixture***10ml
  Heat the above mixed solution at 60℃ for 5 to 10 min and then bubble with N2 gas. And then add the following ingredients.
   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
   Boric acid0.01g
  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.
   Acetic acid17ml
   Propionic acid6ml
   Butyric acid4ml
   iso-Butyric acid1ml
   n-Valeric acid1ml
   iso-Valeric acid1ml
   DL-α-Methylbutyric acid1ml
  ***Vitamin mixture
   Folic acid0.2mg
   p-Aminobenzoic acid20mg
   Choline chloride100mg
   Distilled water1L
  ****3% Cellulose suspension which is ball-milled for 3 days.