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Main sol. 120c

Internal Solution-ID: S217

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.35 g 0.349 2.001
KH2PO4 0.23 g 0.229 1.683
NH4Cl 0.50 g 0.498 9.31
MgSO4 x 7 H2O 0.50 g 0.498 2.021
CaCl2 x 2 H2O 0.25 g 0.249 1.694
NaCl 2.25 g 2.241 38.348
FeSO4 x 7 H2O solution (0.1% w/v) 2.00 ml - -
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID)
2.00 g 1.992 -
Casitone
(BD BBL)
2.00 g 1.992 -
Na-acetate 10.00 g 9.96 121.415
Sucrose 17.10 g 17.032 49.757
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
NaHCO3 2.00 g 1.992 23.713
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.701
Na2S x 9 H2O 0.30 g 0.299 1.244
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, vitamins, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then add and dissolve bicarbonate, adjust pH to 6.8 and 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. Cysteine and sulfide are autoclaved separately under 100% N2 gas atmosphere as concentrated stock solutions. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 6.8 - 7.0, if necessary.
120c METHANOSARCINA MAZEI (GOE1) MEDIUM