National Institute of Technology and Evaluation
NITE Biological Resource Center
Biotechnology Center
HOME > NBRC > NBRC Culture Collection > NBRC Culture Catalogue Search > Catalogue Search Result

Medium Detail Information

Medium No.
1356 
Medium
Medium for haloalkaliphilic methanogen 
Composition
 Mineral base 1*500ml
 Mineral base 2**500ml
 MgCl2·6H2O0.2g
 Bacto Yeast Extract (Difco)0.02g
 Coenzyme M0.014g
 Vitamin solution***5ml
 Trace element solution****10ml
 Methanol2ml
 Sodium formate3.4g
 Sodium acetate0.16g
 FeS slurry solution*****2ml
 Na2S·9H2O0.12g
  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
   Na2CO3185g
   NaHCO335g
   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
   NaCl240g
   NH4Cl0.5g
   K2HPO42.5g
   KCl5g
   Distilled water1L
  Mix ingredients and autoclave under a N2 atmosphere. Final pH should be 7.3.
       
  ***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.
       
  *****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.
 
Comment
 


contact us
Disclaimer Terms of use
National Institute of Technology and Evaluation (JCN 9011005001123)
2-49-10 Nishihara, Shibuya-ku, Tokyo, Japan