| Sodium pyruvate | 2.2 | g | | NH4Cl | 0.54 | g | | KH2PO4 | 0.14 | g | | MgCl2·6H2O | 3 | g | | CaCl2·2H2O | 0.15 | g | | NaCl | 25 | g | | NaHCO3 | 2.5 | g | | Trace element solution* | 2 | ml | | Vitamin solution** | 2 | ml | | Cysteine-HCl | 0.3 | g | | Na2S·9H2O | 0.3 | g | | Distilled water | 1 | L | | | | Mix ingredients except for sodium pyruvate, vitamin solution, cysteine-HCl and Na2S·9H2O, and autoclaved under a N2/CO2 atmosphere (80/20). Concentrated cysteine-HCl and Na2S·9H2O solutions are sterilized separately under a N2 atmosphere in tightly closed vessels. Prior to inoculation, add filter-sterile vitamin and sodium pyruvate solutions, cysteine-HCl, and Na2S·9H2O. Final pH is 7. | | | | | | | | | | *Trace element solution | | | | HCl (35%) | 7 | ml | | | | FeCl3·6H2O | 1500 | mg | | | | H3BO3 | 6 | mg | | | | MnCl2·4H2O | 100 | mg | | | | CoCl2·6H2O | 190 | mg | | | | NiCl2·6H2O | 24 | mg | | | | CuCl2·2H2O | 2 | mg | | | | ZnCl2 | 70 | mg | | | | Na2MoO4·2H2O | 36 | mg | | | | Na2SeO4 | 4 | mg | | | | Na2WO4 | 4 | mg | | | | KAl(SO4)2·12H2O | 20 | mg | | | | Distilled water | 1 | L | | | | | | | | | | **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 | 5 | mg | | | | Distilled water | 1 | L |
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