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Main sol. 1630b

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Internal Solution-ID: S3386

Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 26.00 g 26 444.901
MgSO4 x 7 H2O 5.00 g 5 20.286
MgCl2 x 6 H2O 5.00 g 5 24.594
CaCl2 x 2 H2O 1.50 g 1.5 10.203
KBr 0.10 g 0.1 0.84
KH2PO4
(0.10% w/v) 		2.00 ml 0.002 0.015
NaHCO3 0.25 g 0.25 2.976
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
NH4Cl 0.05 g 0.05 0.935
Na-pyruvate 0.05 g 0.05 0.454
Wolin's vitamin solution 10.00 ml - -
Catalase
(SIGMA C1345) (0.10% w/v) 		2.50 ml 0.003 -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, trace elements, selenite-tungstate, ammonium chloride, pyruvate, vitamins, and catalase) and dispense medium in Erlenmeyer flasks with screw caps (e.g., from SCHOTT), then autoclave. Add bicarbonate, trace elements, selenite-tungstate, ammonium chloride, pyruvate and vitamins from stock solutions sterilized by filtration. Adjust pH of the complete medium to 6.8 using a sterile solution of sodium bicarbonate (10% w/v) or 1 N HCl.
2 After inoculation, add to the medium 5 - 10 units/ml catalase from a freshly prepared 1.00 mg/ml stock solution sterilized by filtration.
3 Note: Use at least 5 - 10% (v/v) inoculum and incubate in the dark without shaking.
1630b NITROSOPUMILUS UREIPHILUS MEDIUM