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Medium Detail Information

Medium No.
1305 
Medium
Medium for natronophilic methylotrophic methanogen 
Composition
 Mineral base 1*700ml
 Mineral base 2**300ml
 MgSO4·7H2O0.25g
 NH4Cl0.2g
 Bacto Yeast Extract (Difco)0.02g
 Coenzyme M0.14g
 Vitamin solution***10ml
 Trace element solution****10ml
 Methanol3ml
 Na2S·9H2O0.25g
  Mix mineral base 1 with mineral base 2 in 7:3 propotion in tightly closed vessels under a N2 atmosphere. Separately autoclave concentrated MgSO4·7H2O, NH4Cl, Bacto Yeast Extract, coenzyme M and Na2S·9H2O under a N2 atmosphere in tightly closed vessels. Add the concentrated solutions, and filter-sterile vitamin solution, trace element solution and methanol. 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 10.
       
  *Mineral base 1
   Na2CO395g
   NaHCO315g
   NaCl16g
   K2HPO41g
   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
   Na2CO315g
   NaHCO320g
   NaCl3g
   K2HPO41g
   Distilled water1L
  Mix ingredients, adjust pH to 9.5 with HCl and autoclave under a N2 atmosphere. Final pH should be 9.5.
       
  ***Vitamin solution
   Biotin2mg
   Folic acid2mg
   Pyridoxine-HCl10mg
   Thiamine-HCl5mg
   Riboflavin5mg
   Nicotinic acid5mg
   Ca-pantothenate5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L
       
  ****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
   Distilled water1L
  First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
 
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