Main sol. 924b
Internal Solution-ID: S1901
Equipment needed: Autoclave
Hungate tubes
Filter
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 1.00 | g | 0.989 | 16.925 | |
| MgCl2 x 6 H2O | 1.00 | g | 0.989 | 4.865 | |
| KCl | 0.50 | g | 0.495 | 6.634 | |
| CaCl2 x 2 H2O | 0.40 | g | 0.396 | 2.691 | |
| K2HPO4 | 0.40 | g | 0.396 | 2.272 | |
| NH4Cl | 1.00 | g | 0.989 | 18.492 | |
| Modified Wolin's mineral solution | 10.00 | ml | - | - | |
|
Yeast extract
(OXOID) |
2.00 | g | 1.978 | - | |
|
Trypticase peptone
(BD BBL) |
2.00 | g | 1.978 | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
| NaHCO3 | 4.00 | g | 3.956 | 47.097 | |
| Na-formate | 6.80 | g | 6.726 | 98.902 | |
| Na-acetate | 1.60 | g | 1.583 | 19.292 | |
| Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.25 | g | 0.247 | 1.408 | |
| Na2S x 9 H2O | 0.25 | g | 0.247 | 1.03 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients (except bicarbonate, formate, acetate vitamins, and reducing agents), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add solid bicarbonate, adjust pH to 6.8 - 7.0, dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add formate, acetate, vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The pH of the complete medium should be 7.2. | |||||
Media with this solution
| 924b | METHANOSARCINA MB MEDIUM (1 g/l NaCl) |