| | Sodium acetate | 1.7 | g |
| | NH4Cl | 1 | g |
| | KH2PO4 | 0.3 | g |
| | NaCl | 0.6 | g |
| | MgCl2·6H2O | 0.1 | g |
| | CaCl2·2H2O | 0.08 | g |
| | Trace elements solution* | 10 | ml |
| | Vitamin solution** | 10 | ml |
| | Bacto Yeast Extract (Difco) | 0.5 | g |
| | Hipolypepton (Nihon seiyaku) | 0.5 | g |
| | Resazurin | 1 | mg |
| | KHCO3 | 4.5 | g |
| | Cysteine-HCl | 0.3 | g |
| | Na2S·9H2O | 0.3 | g |
| | Distilled water | 1 | L |
|
| | | Mix ingredients except for vitamin solution, cysteine-HCl and Na2S·9H2O, and autoclave under N2/CO2 (80/20) condition. Separately autoclave cysteine-HCl and Na2S·9H2O as a 3% solution under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin, cysteine-HCl and Na2S·9H2O solutions. Final pH is 7.2-7.5. |
| | | | | | | |
| | | *Trace elements solution |
| | | | Nitrilotriacetic acid (NTA) | 4.1 | g |
| | | | FeCl3·6H2O | 0.36 | g |
| | | | MnCl2·4H2O | 0.09 | g |
| | | | CoCl2·6H2O | 0.15 | g |
| | | | CaCl2·2H2O | 0.018 | g |
| | | | ZnCl2 | 0.09 | g |
| | | | H3BO3 | 0.017 | g |
| | | | Na2MoO4·2H2O | 0.009 | g |
| | | | NiCl2·6H2O | 0.018 | g |
| | | | Na2SeO3 | 1.6 | mg |
| | | | Na2WO4 | 1.6 | mg |
| | | | Distilled water | 1 | L |
|
| | | First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add other minerals. Final pH 7.0. |
| | | | | | | |
| | | **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 |