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Medium No.	1445
Medium
Composition
Na2SO4 2.84 g   NH4Cl 0.27 g   KH2PO4 0.2 g   MgCl2·6H2O 0.4 g   CaCl2·2H2O 0.15 g   NaCl 1.17 g   KCl 0.3 g   Trace element solution* 1 ml   Selenite-tungstate solution** 1 ml   Yeast extract 1 g   Resazurin 1 mg   NaHCO3 4.5 g   Sodium benzoate 0.6 g   Vitamin solution*** 10 ml   Na2S·9H2O 0.4 g   Distilled water 990 ml     Mix ingredients (except NaHCO3, sodium benzoate, vitamin solution and Na2S·9H2O) and dispense into suitable culture vessels under a stream of N2/CO2 (80/20) gas, seal with butyl rubber stoppers, and then autoclave. Separately autoclave NaHCO3 under a N2/CO2 (80/20) atmosphere. Separately autoclave Na2S·9H2O under a N2 atmosphere. Sterilize sodium benzoate and vitamin solution by filtration. To complete the medium, add NaHCO3, sodium benzoate, vitamin solution and Na2S·9H2O. Adjust pH of the complete medium to 7.0-7.2, if necessary.                   *Trace element solution       HCl (35%) 7 ml       FeCl3·6H2O 1.5 g       ZnCl2 70 mg       MnCl2·4H2O 100 mg       H3BO3 6 mg       CoCl2·6H2O 190 mg       CuCl2·2H2O 2 mg       NiCl2·6H2O 24 mg       Na2MoO4·2H2O 36 mg       Distilled water 1 L     First dissolve FeCl3 in conc. HCl, and dilute in water, then add the other minerals.                   **Selenite-tungstate solution       NaOH 0.5 g       Na2SeO3·5H2O 3 mg       Na2WO4·2H2O 4 mg       Distilled water 1 L                   ***Vitamin solution       Biotin 2 mg       Folic acid 2 mg       Pyridoxine-HCl (VitaminB6) 10 mg       Thiamine-HCl·H2O (Vitamin B1) 5 mg       Riboflavin (Vitamine B2) 5 mg       Nicotinic acid 5 mg       D-Ca-pantothenate 5 mg       Vitamin B12 0.1 mg       p-Aminobenzoic acid 5 mg       Lipoic acid 5 mg       Distilled water 1 L     Adjust pH to 7.0 with NaOH.
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