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Medium Detail Information

Medium No.
Medium for haloalkaliphilic methanogen 
 Mineral base 1*500ml
 Mineral base 2**500ml
 Bacto Yeast Extract (Difco)0.02g
 Coenzyme M0.014g
 Vitamin solution***5ml
 Trace element solution****10ml
 Sodium formate3.4g
 Sodium acetate0.16g
 FeS slurry solution*****2ml
  Mix mineral base 1 with mineral base 2 in 1:1 proportion in tightly closed vessels under a N2 atmosphere. Separately autoclave concentrated Bacto Yeast Extract, coenzyme M, sodium acetate, Na2S·9H2O and FeS solution under a N2 atmosphere in tightly closed vessels. Add the autoclaved concentrated solutions, and filter-sterile concentrated MgCl2·6H2O, vitamin solution, trace element solution, methanol and sodium formate solution. After re-exchanging with N2, add 1 drop of filter-sterile 10% dithionaite solution in 1 M NaHCO3 by syringe. Final pH of the complete medium should be 9.
  *Mineral base 1
   Distilled water1L
  Mix ingredients and autoclave under a N2 atmosphere. Because some precipitate appears on the bottom for 1 day after autoclaving, use only supernatant for making the medium. Final pH should be 10.
  **Mineral base 2
   Distilled water1L
  Mix ingredients and autoclave under a N2 atmosphere. Final pH should be 7.3.
  ***Vitamin solution
   Folic acid2mg
   Nicotinic acid5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L
  ****Trace elements solution
   Nitrilotriacetic acid (NTA)12.8g
   Distilled water1L
  First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
  *****FeS slurry solution
  Mix 0.5 M FeSO4·7H2O and 0.5 M Na2S·9H2O solutions, wash with pure water several times, finally resuspend the FeS precipitate as 20% solution and autoclave (110°C, 30 min) under a N2 atmosphere.

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