| Sodium lactate | 1.1 | g | | Disodium fumarate | 1.6 | g | | KH2PO4 | 0.225 | g | | K2HPO4 | 0.225 | g | | NaCl | 0.46 | g | | (NH4)2SO4 | 0.225 | g | | MgSO4·7H2O | 0.117 | g | | CaCl2·2H2O | 0.14 | g | | Bacto Yeast Extract (Difco) | 1 | g | | Na2S2O3·5H2O | 2.5 | g | | Trace element mixture* | 2 | ml | | Vitamin solution** | 10 | ml | | NaHCO3 | 4.2 | g | | Resazurin | 0.5 | mg | | Na2S·9H2O | 0.25 | g | | Cysteine-HCl | 0.25 | g | | Distilled water | 1 | L | | | | Mix ingredients except for lactate, fumarate, vitamin, Na2S·9H2O and cysteine-HCl, and autoclave at 121°C for 20 min under a N2/CO2 (80/20) atmosphere. Concentrated Na2S·9H2O and cysteine-HCl solutions are sterilized separately under a N2 atmosphere in tightly closed vessels. Add filter-sterile lactate, fumarate and vitamin solutions, Na2S·9H2O, and cysteine-HCl. Final pH is 6.8-7.2. | | | | | | | | | | *Trace element mixture | | | | 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 | | | | Distilled water | 1 | L | | | | First dissolve FeCl3 in concentrated HCl, and dilute in water, then add the other minerals. | | | | | | | | | | **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 | | | | Vitamin B12 | 0.1 | mg | | | | p-Aminobenzoic acid | 5 | mg | | | | Lipoic acid | 5 | mg | | | | Distilled water | 1 | L |
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