| | KH2PO4 | 0.2 | g |
| | NH4Cl | 0.06 | g |
| | Cysteine-HCl | 0.05 | g |
| | Ferric quinate solution* | 2 | ml |
| | Wolfe's vitamin solution** | 4 | ml |
| | Wolfe's mineral solution*** | 2 | ml |
| | Sodium fumarate | 0.58 | g |
| | Sodium pyruvate | 0.44 | g |
| | Distilled water | 1 | L |
| | pH 7 | |
|
| | | Mix ingredients except for vitamin solution and cysteine-HCl. Dispense the medium into vials suitable for anaerobic cultures, sparge with argon gas to establish anoxic condition (at least 30 min), seal with butyl rubber stoppers, and autoclave at 121°C for 15 min. Separately autoclave cysteine-HCl (0.5% solution) under argon atmosphere. Sterilize vitamin solution by filtration. Prior to inoculation, aseptically and anaerobically add the vitamin and cysteine-HCl solutions to the medium, and sparge the vials with sterile argon gas for 5 min. The cysteine-HCl more than the above-mentioned amount (0.05 g/L) is unsuitable for growth of strain RS-1. And the strain produces only a small amount of magnetite in the presence of sulfate. |
| | | | | | | |
| | | *Ferric quinate solution |
| | | | FeCl3 | 0.27 | g |
| | | | Quinate | 0.19 | g |
| | | | Distilled water | 100 | ml |
| | | | | | | |
| | | **Wolfe's 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 |
| | | | | | | |
| | | ***Wolfe's mineral solution |
| | | | Nitrilotriacetic acid (NTA) | 1.5 | g |
| | | | MgSO4·7H2O | 3 | g |
| | | | MnSO4·H2O | 0.5 | g |
| | | | NaCl | 1 | g |
| | | | FeSO4·7H2O | 0.1 | g |
| | | | CoCl2·6H2O | 0.1 | g |
| | | | CaCl2 | 0.1 | g |
| | | | ZnSO4·7H2O | 0.1 | g |
| | | | CuSO4·5H2O | 0.01 | g |
| | | | KAl(SO4)2·12H2O | 0.01 | g |
| | | | H3BO3 | 0.01 | g |
| | | | Na2MoO4·2H2O | 0.01 | g |
| | | | Distilled water | 1 | L |
| | | Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1 L with remaining water and add remaining compounds one at a time. |