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Main sol. 975

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Internal Solution-ID: S2000

Equipment needed: Autoclave Anaerobic workbench Filter Hungate tubes
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.33 6.169
MgCl2 x 6 H2O 0.33 g 0.33 1.623
CaCl2 x 2 H2O 0.33 g 0.33 2.245
KCl 0.33 g 0.33 4.427
KNO3 1.00 g 1 9.891
NaCl 30.00 g 30 513.347
HEPES
(10mM) 		2.38 g 2.38 9.987
Yeast extract 0.20 g 0.2 -
Tryptone 1.00 g 1 14.069
Sucrose 2.00 g 2 5.843
Wolin's vitamin solution 1.00 ml - -
Modified Wolin's mineral solution 1.00 ml - -
Distilled water 1000.00 ml - -
1 Prepare the medium anaerobically, under nitrogen, omitting the CaCl2, MgCl2, KNO3, tryptone, yeast extract, vitamins and sucrose. The pH should be 7.0-7.5. Dispense the medium into vessels suitable for anaerobic growth (Hungate tubes or serum bottles) under an atmosphere of nitrogen and autoclave. To the sterile, cooled medium add, from sterile stock solutions the CaCl2, MgCl2, KNO3, tryptone, yeast extract, vitamins and sucrose. The CaCl2, MgCl2, KNO3, tryptone, yeast extract, and sucrose stock solutions should be autoclaved, while the vitamin solution is sterile filtered. This strain grows anaerobically in the presence of nitrate, but microaerophilic growth has been reported provided the oxygen concentration does not exceed 6 % (v/v). Under anaerobic conditions a redox agent, such as sulphide is not required.
975 OCEANITHERMUS PROFUNDUS MEDIUM