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Main sol. J1186

Internal Solution-ID: S5299

Equipment needed: Autoclave Anaerobic workbench Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.10 g 0.096 0.706
MgCl2 x 6 H2O 4.00 g 3.842 18.9
CaCl2 x 2 H2O 1.00 g 0.961 6.534
NH4Cl 0.50 g 0.48 8.979
NaCl 25.00 g 24.015 410.941
Yeast extract 0.10 g 0.096 -
Sodium formate 0.68 g 0.653 9.605
Sodium acetate 0.08 g 0.077 0.937
Trace vitamins solution 2.00 ml - -
Trace mineral solution 1.00 ml - -
Selenite-tungstate solution
(Medium No. 852)
1.00 ml - -
Resazurin 1.00 mg 9.6e-4 0.004
Distilled water 1000.00 ml - -
1 Mix components and adjust pH to 7.5. Bring to a boil and cool down under a H2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels (e.g., 20 ml medium in 60 ml serum bottles) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobically add per liter the following solutions (autoclaved or *filter-sterilized):
NaHCO3
(*, 8%)
25.00 ml - -
L-Cysteine HCl x H2O
(5%)
6.00 ml - -
Na2S x 9 H2O
(5%)
6.00 ml - -
2 Pressurize the inoculated vessels to 100 kPa H2-CO2 (4:1, v/v)
J1186 MARINE METHANOBACTERIUM MEDIUM