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Medium Detail Information

Medium No.	1446
Medium
Composition
Na2SO4 3 g   NH4Cl 0.3 g   KH2PO4 0.2 g   MgCl2·6H2O 0.4 g   CaCl2·2H2O 0.15 g   NaCl 1 g   Trace element solution* 1 ml   Resazurin 1 mg   NaHCO3 1 g   Ethanol 1 ml   Sodium benzoate 0.7 g   Rumen fluid** 1 ml   Vitamin solution*** 10 ml   Na2S·9H2O 50 mg   Sodium dithionite 25 mg   Distilled water 990 ml     Mix ingredients (except NaHCO3, ethanol, sodium benzoate, rumen fluid, vitamin solution, Na2S·9H2O and sodium dithionite) 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 rumen fluid and Na2S·9H2O under a N2 atmosphere. Sterilize sodium benzoate, vitamin solution and sodium dithionite by filtration. To complete the medium, add NaHCO3, ethanol, sodium benzoate, rumen fluid, vitamin solution, Na2S·9H2O and sodium dithionite. Adjust pH of the complete medium to around 7.5, 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.                   **Rumen fluid     Remove solid particles from a fresh rumen fluid by filtration and centrifugation, and then autoclave under a N2 atmosphere. Stock at 4°C.                   ***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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