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Main sol. J492

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Internal Solution-ID: S4250

Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Modified MJ synthetic seawater 1000.0 ml - -
Trace vitamins 10.0 ml - -
NaHCO3 1.0 g 0.99 11.786
Sulfur
(powder) 		3.0 g 2.97 92.619
Resazurin 1.0 mg 9.9e-4 0.004
Na2S x 9 H2O 0.5 g 0.495 2.061
1 Mix ingredients except NaHCO3, sulfur and Na2S x 9H2O, adjust pH to 7.0 with NaOH and autoclave under a N2 atmosphere. Filter-sterilize 8% NaHCO3 solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) containing sulfur under a stream of a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Prior to inoculation, add 5% Na2S x 9H2O (autoclaved and stocked under a N2 atmosphere) to the medium. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).
J492 MMJ-TL MEDIUM