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Main sol. 1630a

Internal Solution-ID: S3384

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
(0.1% w/v)
2.00 ml 0.002 0.015
HEPES buffer solution 10.00 ml - -
NH4Cl 0.05 g 0.05 0.935
Trace element solution SL-11 1.00 ml - -
Na-pyruvate 0.05 g 0.05 0.454
NaHCO3 0.25 g 0.25 2.976
Wolin's vitamin solution 10.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except trace elements, HEPES buffer, ammonium chloride, pyruvate, bicarbonate and vitamins), dispense medium into Erlenmeyr flasks with screw caps (e.g., 20 ml medium per 100 ml flask) and autoclave. Add HEPES buffer and ammonium chloride from stock solutions sterilized by autoclaving. Trace elements, pyruvate, bicarbonate and vitamins are added from stock solutions sterilized by filtration. Adjust pH of the complete medium to 7.2 using sterile solutions of bicarbonate (5% w/v) or 1 N HCl.
2 Note: Use at least 5% (v/v) inoculum and incubate in the dark without shaking.
1630a NITROSOPUMILUS MEDIUM