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Medium Detail Information

Medium No.	1241
Medium	Alkaliphilic spirochaete medium
Composition
NaCl 10 g   K2HPO4 0.2 g   NH4Cl 1 g   KCl 0.2 g   MgSO4·7H2O 0.003 g   Na2S·9H2O 1 g   Yeast extract 0.5 g   Sucrose 5 g   Vitamin solution* 10 ml   Trace element solution** 2 ml   Bicarbonate solution*** 200 ml   Distilled water 800 ml   pH 9.7       Mix ingredients except for sulfide, vitamin and bicarbonate. Dispense the medium into suitable culture vessels under a stream of nitrogen gas, seal with butyl rubber stoppers, and autoclave at 121°C for 20 min. Separately autoclave sulfide solution under a N2 atmosphere. Sterilize vitamin and bicarbonate solutions by filtration. Prior to inoculation, aseptically and anaerobically add the vitamin, bicarbonate and sulfide solutions to the medium.                   *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       Vitamin B12 0.1 mg       p-Aminobenzoic acid 5 mg       Lipoic acid 5 mg       Distilled water 1 L                   **Trace element 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       KAl(SO4)2·12H2O 0.02 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.                   ***Bicarbonate solution       Na2CO3 50 g       NaHCO3 75 g       Distilled water 1 L     Sterilized by filtration.
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