Main sol. 945
Internal Solution-ID: S1956
Equipment needed: Hungate tubes
Gassing station
Water bath
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NH4Cl | 1.00 | g | 1 | 18.695 | |
| MgCl2 x 6 H2O | 0.20 | g | 0.2 | 0.984 | |
| CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.68 | |
| KCl | 0.10 | g | 0.1 | 1.341 | |
| NaCl | 30.00 | g | 30 | 513.347 | |
| Na-acetate | 0.83 | g | 0.83 | 10.118 | |
|
MES
(SIGMA) |
1.95 | g | 1.95 | 9.988 | |
| Yeast extract | 5.00 | g | 5 | - | |
|
Trypticase peptone
(BD BBL) |
5.00 | g | 5 | - | |
| K2HPO4 | 0.30 | g | 0.3 | 1.722 | |
| KH2PO4 | 0.30 | g | 0.3 | 2.205 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
|
Sulfur
(powdered) |
10.00 | g | 10 | 311.818 | |
| L-Cysteine HCl x H2O | 0.30 | g | 0.3 | 1.708 | |
| Na2S x 9 H2O | 0.30 | g | 0.3 | 1.249 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients except sulfur, cysteine and sulfide. Sparge medium with 100% N2 gas for at least 30 - 45 min to make it anoxic and adjust pH to 6.0 with NaOH. Distribute the medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur. For sterilization heat the cultivation vessels containing medium plus sulfur in boiling water for 2 - 3 hours on each of 3 successive days! Prior to inoculation add cysteine and sulfide and from sterile anoxic stock solutions prepared under 100% N2 gas. | |||||
| 2 After inoculation pressurize tubes to 2 bar overpressure with sterile 100% N2 gas. | |||||
Media with this solution
| 945 | MARINITOGA PIEZOPHILA MEDIUM |