Main sol. J944
Internal Solution-ID: S4962
Equipment needed: Autoclave
Anaerobic workbench
Filter
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 1.00 | g | 0.988 | 16.909 | |
| KCl | 0.50 | g | 0.494 | 6.627 | |
| MgCl2 x 6 H2O | 0.40 | g | 0.395 | 1.944 | |
| NH4Cl | 0.25 | g | 0.247 | 4.618 | |
| CaCl2 x 2 H2O | 0.15 | g | 0.148 | 1.008 | |
| Trace elements solution | 10.00 | ml | - | - | |
| Resazurin | 1.00 | mg | 9.9e-4 | 0.004 | |
| Distilled water | 946.00 | ml | - | - | |
| 1 Mix components thoroughly, and autoclave under a N2-CO2 (80:20, v/v) gas atmosphere. After cooling, aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized): | |||||
|
KH2PO4
(10%) |
2.00 | ml | - | - | |
|
NaHCO3
(*, 8%) |
32.00 | ml | - | - | |
|
Vitamin solution
(*) |
10.00 | ml | - | - | |
| 2 Aseptically distribute the medium into culture vessels under a H2-CO2 (80:20, v/v) gas mixture and seal with butyl rubber stoppers (leave >80% gas phase by volume). Finally, add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N2 atmosphere): | |||||
|
L-Cysteine HCl x H2O
(5%) |
6.00 | ml | - | - | |
|
Na2S x 9 H2O
(5%) |
6.00 | ml | - | - | |
| 3 Pressurize the inoculated vessels to 200 kPa H2-CO2 (80:20, v/v). | |||||
Media with this solution
| J944 | MW MEDIUM FOR METHANOSARCINA |