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Medium No.	1433
Medium	Modified WS medium 2
Composition
KH2PO4 0.136 g   NH4Cl 0.54 g   MgCl2·6H2O 3.05 g   CaCl2·2H2O 0.147 g   NaCl 20 g   Methanol 0.8 ml   Coenzyme M 0.0014 g   NaHCO3 2.52 g   Vitamin solution* 2 ml   Trace element solution** 1 ml   Resazurin 1 mg   Cysteine-HCl 0.3 g   Na2S·9H2O 0.3 g   Distilled water 1 L     Mix ingredients except for methanol, vitamin solution, cysteine-HCl and Na2S·9H2O, and autoclave under a N2/CO2 atmosphere (80/20). Separately autoclave cysteine-HCl and Na2S·9H2O solutions as 3% solutions under a N2 atmosphere. Sterilize methanol and vitamin solution by filtration. Prior to inoculation, add methanol, vitamin solution, cysteine-HCl and Na2S·9H2O.                   *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       p-Aminobenzoic acid 1 mg       Vitamin B12 0.01 mg       Distilled water 1 L                   **Trace elements 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       Distilled water 1 L     First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
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