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Medium No.
Medium for Co-culture of Strain JT with Methanospirillum hungatei NBRC 100397
 Yeast extract0.2g
 Vitamin solution*3ml
 Trace element solution**1ml
 Se/W solution***1ml
 Resazurin solution****1ml
 Distilled water1L
  This medium is for co-culture of strain JT with Methanospirillum hungatei strain NBRC 100397.
  For vial cultures, dissolve all ingredients except for Na-terephthalate, Na2S·9H2O and cysteine-HCl (up to 900ml), flush N2/CO2 (80/20), and autoclave. Add 0.5 ml of each of filter-sterilized Na2S·9H2O solution (0.6g Na2S·9H2O/50 ml) and cysteine-HCl solution (0.6g cysteine-HCl/ 50ml) per each vial separately. Add 0.2 ml of neutralized and filter-sterilized 0.2 M Na-terephthalate solution. Yeast extract is also added at final concentration about 0.02%. Inoculum size 10-20%. Growth was observed after 3-4 weeks of incubation.
  *Vitamin solution
   Folic acid0.9mg
   Nicotinic acid0.2mg
   Vitamin B122.7mg
   p-Aminobenzoic acid0.3mg
   Lipoic acid0.4mg
   Distilled water1L
  **Trace element solution
   Nitrilotriacetic acid (NTA)12.8g
   Distilled water1L
  First dissolve NTA in 200 ml of water and adjust the pH to 6.5 with KOH.
  ***Se/W solution
  ****Resazurin solution
   Distilled water1L