| KCl | 0.34 | g | | MgCl2·6H2O | 4 | g | | MgSO4·7H2O | 3.45 | g | | NH4Cl | 0.25 | g | | CaCl2·2H2O | 0.14 | g | | K2HPO4 | 0.14 | g | | NaCl | 18 | g | | NaHCO3 | 3 | g | | Bacto Yeast Extract (Difco) | 0.5 | g | | Sodium lactate | 1.5 | g | | Fe(NH4)2(SO4)2·7H2O | 2 | mg | | Trace element solution* | 10 | ml | | Resazurin | 1 | mg | | Na2S·9H2O | 0.5 | g | | Distilled water | 1 | L | | | Dissolve ingredients except bicarbonate and sulfide, bring to boil for few minutes, then cool quickly to room temperature while gassing with N2/CO2 (80/20), add sodium bicarbonate and adjust pH to 6.9. Distribute in serum bottles (20 ml per 100 ml bottle) under N2/CO2, seal, pressurize bottles up to 2 bar overpressure, then autoclave. Before use, reduce the medium with sodium sulfide from anaerobic sterile neutral stock solution. | | | | | | | | | | *Trace element solution | | | | Nitrilotriacetic acid | 1.5 | g | | | | MgSO4·7H2O | 3 | g | | | | MnSO4·2H2O | 0.5 | g | | | | NaCl | 1 | g | | | | FeSO4·7H2O | 0.1 | g | | | | CoSO4·7H2O | 0.18 | g | | | | CaCl2·2H2O | 0.1 | g | | | | ZnSO4·7H2O | 0.18 | g | | | | CuSO4·5H2O | 0.01 | g | | | | KAl(SO4)2·12H2O | 4 | mg | | | | H3BO3 | 0.01 | g | | | | Na2MoO4·2H2O | 0.01 | g | | | | NiCl2·6H2O | 0.025 | g | | | | Na2SeO3·5H2O | 0.8 | mg | | | | Na2WO4·H2O | 0.8 | mg | | | | Distilled water | 1 | L | | | | First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final pH 7.0 (with KOH). |
|