Main sol. 120d
Internal Solution-ID: S218
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
K2HPO4 | 0.35 | g | 0.342 | 1.962 | |
KH2PO4 | 0.23 | g | 0.225 | 1.651 | |
NH4Cl | 0.50 | g | 0.488 | 9.128 | |
MgCl2 x 6 H2O | 10.00 | g | 9.766 | 48.035 | |
CaCl2 x 2 H2O | 0.15 | g | 0.146 | 0.996 | |
NaCl | 23.00 | g | 22.461 | 384.342 | |
FeSO4 x 7 H2O solution (0.1% w/v) | 2.00 | ml | - | - | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
Na2CO3 | 1.00 | g | 0.977 | 9.214 | |
Methanol
(50% v/v) |
20.00 | ml | 15.469 | 482.765 | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
L-Cysteine HCl x H2O | 0.30 | g | 0.293 | 1.668 | |
Na2S x 9 H2O | 0.30 | g | 0.293 | 1.22 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except carbonate, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N2 gas atmosphere as concentrated solutions in tightly closed tubes. Carbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved separately. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 6.8 - 7.0, if necessary. |
120d | METHANOSARCINA MARINE MEDIUM |