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Medium No.	1443
Medium
Composition
Na2SO4 2 g   NH4Cl 1 g   KH2PO4 0.5 g   MgSO4·7H2O 1 g   CaCl2·2H2O 0.1 g   Trace element solution* 1 ml   Selenite-tungstate solution** 1 ml   Yeast extract 1 g   Resazurin 1 mg   Na2S2O3·5H2O 2 g   NaHCO3 2 g   Vitamin solution*** 10 ml   Sodium pyruvate 2.5 g   Na2S·9H2O 0.1 g   Distilled water 990 ml     Mix ingredients (except Na2S2O3·5H2O, NaHCO3, vitamin solution, sodium pyruvate, Na2S·9H2O and sodium dithionate) and dispense into suitable culture vessels under a stream of N2 gas, seal with butyl rubber stoppers, and then autoclave. Separately autoclave NaHCO3 under a N2/CO2 (80/20) atmosphere, and Na2S·9H2O under a N2 atmosphere. Sterilize Na2S2O3·5H2O, vitamin solution and sodium pyruvate by filtration. To complete the medium, add Na2S2O3·5H2O, NaHCO3, vitamin solution, sodium pyruvate and Na2S·9H2O. Adjust pH of the complete medium to around 7.5, if necessary.     Addition of 10-20 mg/L sodium dithionite may stimulate growth at the begining.                   *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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