| | KNO3 | 1 | g |
| | MgSO4·6H2O | 1 | g |
| | CaCl2·2H2O | 0.26 | g |
| | Ferric EDTA solution* | 0.1 | ml |
| | Molybdate solution** | 1 | ml |
| | Trace elements solution*** | 2 | ml |
| | 0.4 M Phosphate buffer **** | 10 | ml |
| | Artificial seawater | 1 | L |
| | pH 6.8 | |
|
| | | Mix ingredients except for phosphate buffer, adjust pH to 6.8, and dispense the medium into suitable culture vessels, seal with butyl rubber stoppers, and autoclave at 121°C for 20 min. After autoclaved, aseptically add phosphate buffer which is separately autoclaved. Prior to inoculation, aseptically add the filter-sterilized methane gas to the vessel. The methane and air ratio in the headspace is about 1:4. Methane should be of high purity. |
| | | | | | | |
| | | *Ferric EDTA solution |
| | | | Fe(III)-EDTA | 38 | g |
| | | | Distilled water | 1 | L |
| | | | | | | |
| | | **Molybdate solution |
| | | | Na2MoO4·2H2O | 0.26 | g |
| | | | Distilled water | 1 | L |
| | | | | | | |
| | | ***Trace elements solution |
| | | | CuSO4·5H2O | 0.2 | g |
| | | | FeSO4·7H2O | 0.5 | g |
| | | | ZnSO4·7H2O | 0.4 | g |
| | | | H3BO3 | 0.015 | g |
| | | | CoCl2·6H2O | 0.05 | g |
| | | | 2Na-EDTA | 0.25 | g |
| | | | MnCl2·4H2O | 0.02 | g |
| | | | NiCl2·6H2O | 0.01 | g |
| | | | Distilled water | 1 | L |
|
| | | Dissolve the above in the specified order in distilled water and dilute to 1 litre. Store in the dark. |
| | | | | | | |
| | | ****0.4 M Phosphate buffer: |
|
| | | Mix 0.4 M Na2HPO4 and 0.4 M KH2PO4 solutions, and adjust pH to 6.8 (approximately volume rate is 2:1). |