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Medium Detail Information

Medium No.	1305
Medium	Medium for natronophilic methylotrophic methanogen
Composition
Mineral base 1* 700 ml   Mineral base 2** 300 ml   MgSO4·7H2O 0.25 g   NH4Cl 0.2 g   Bacto Yeast Extract (Difco) 0.02 g   Coenzyme M 0.14 g   Vitamin solution*** 10 ml   Trace element solution**** 10 ml   Methanol 3 ml   Na2S·9H2O 0.25 g     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       Na2CO3 95 g       NaHCO3 15 g       NaCl 16 g       K2HPO4 1 g       Distilled water 1 L     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       Na2CO3 15 g       NaHCO3 20 g       NaCl 3 g       K2HPO4 1 g       Distilled water 1 L     Mix ingredients, adjust pH to 9.5 with HCl and autoclave under a N2 atmosphere. Final pH should be 9.5.                   ***Vitamin solution       Biotin 2 mg       Folic acid 2 mg       Pyridoxine-HCl 10 mg       Thiamine-HCl 5 mg       Riboflavin 5 mg       Nicotinic acid 5 mg       Ca-pantothenate 5 mg       p-Aminobenzoic acid 1 mg       Vitamin B12 0.01 mg       Distilled water 1 L                   ****Trace elements solution       Nitrilotriacetic acid (NTA) 12.8 g       FeCl3·6H2O 1.35 g       MnCl2·4H2O 0.1 g       CoCl2·6H2O 0.024 g       CaCl2·2H2O 0.1 g       ZnCl2 0.1 g       CuCl2·2H2O 0.025 g       H3BO3 0.01 g       Na2MoO4·2H2O 0.024 g       NaCl 1 g       NiCl2·6H2O 0.12 g       Na2SeO4 0.004 g       Na2WO4 0.004 g       Distilled water 1 L     First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
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