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Medium Detail Information

Medium No.	404
Medium	Oceanithermus profundus Medium
Composition
NH4Cl 0.33 g   MgCl2·6H2O 0.33 g   CaCl2·2H2O 0.33 g   KCl 0.33 g   KNO3 0.33 g   NaCl 30 g   HEPES 2.38 g   Yeast extract 0.2 g   Tryptone 1 g   Sucrose 2 g   Vitamin solution* 1 ml   Trace element solution** 1 ml   Distilled water 1 L     Prepare the medium anaerobically, under N2, omitting the CaCl2, MgCl2, KNO3, tryptone, yeast extract, vitamins and sucrose. The pH should be 7.0-7.5. Dispense the medium into vessels suitable for anaerobic growth (Hungate tubes or serum bottles) under an atmosphere of nitrogen and autoclave. To the sterile, cooled medium add, from sterile stock solutions the CaCl2, MgCl2, KNO3, tryptone, yeast extract, vitamins and sucrose. The CaCl2, MgCl2, KNO3, tryptone, yeast extract, and sucrose stock solutions should be autoclaved, while the vitamin solution is filter-sterilized.                   *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 1.5 g       MgSO4·7H2O 3 g       MnSO4·2H2O 0.5 g       NaCl 1 g       FeSO4·7H2O 0.1 g       CoSO4·7H2O 0.18 g       CaCl2·2H2O 0.1 g       ZnSO4·7H2O 0.18 g       CuSO4·5H2O 0.01 g       KAl(SO4)2·12H2O 0.02 g       H3BO3 0.01 g       Na2MoO4·2H2O 0.01 g       NiCl2·6H2O 0.025 g       Na2SeO3·5H2O 0.3 mg       Distilled water 1 L     First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final pH 7.0 (with KOH).
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