SDM culture medium
From Global Methane Hub Wikipedia
- Preparation of SDM culture medium:
Mineral Sol. 1 (1L)
| K2HPO4 | 6 g |
Mineral Sol. 2 (1L)
| NaCl | 12 g |
| KH2PO4 | 6 g |
| CaCl2・2H2O | 1.2 g |
| MgSO4・7H2O | 1.2 g |
VFA Sol. (per liter)
| Acetic acid | 13.7 ml |
| Propionic acid | 6 ml |
| Butyric acid | 3.68 ml |
| Isobutyric acid | 1.1 ml |
| n-valeric acid | 1.1 ml |
| isovaleric acids | 1.1 ml |
| 2-methylbutyric acid | 0.94 ml |
| Phenylacetic acid (25uM) | 3.4 mg |
| Phenylpropionic acid (25uM) | 3.75 mg |
| 0.2N NaOH | 700 ml |
*Adjust pH7.5 and adjust volume to 1L
Cysteine Sulfide Sol. (200ml)
| NaOH | 1.25 g |
| Cysteine-HCl | 5 g |
| Na2S.9H2O | 5 g |
| ddH2O | 200 ml |
- Dissolve NaOH in H2O, then boil under N2,
- Cool to RT and add Cysteine-HCl and Na2S.9H2O
Note: Cystine-HCl (0.5 g/L) and sodium sulfide (0.5 g/L) can be added directly to the medium.
Hemin Sol. (100ml)
| Hemin | 50 mg |
| 1N NaOH | 1 ml |
- Make to 100 ml with ddH2O
- Autoclave
Final mixture (1L)
| Mineral Sol. 1 | 50 ml |
| Mineral Sol. 2 | 50 ml |
| Resazurin (0.1%) | 1 ml |
| Glucose | 4 g |
| (NH4)2SO4 for N source | 4 g |
| Hemin Sol. (0.1%) | 0.5 ml |
| Trypticase | 2 g |
| Yeast extract | 0.5 g |
Boiling and cool down with CO2 gas
Then add inside the anaerobic chamber
| VFA sol. | 10 ml |
| NaHCO3 | 4 g |
| Cysteine sulfide Sol. | 20 ml |
- Adjust pH 6.7 to 6.8 inside the chamber
- Move bottle into anaerobic chamber, aliquoting medium
- Autoclave
