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Main sol. 1630

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Internal Solution-ID: S3382

Equipment needed: Filter Autoclave Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 1 17.112
MgCl2 x 6 H2O 0.40 g 0.4 1.968
CaCl2 x 2 H2O 0.10 g 0.1 0.68
KH2PO4 0.20 g 0.2 1.47
KCl 0.50 g 0.5 6.707
Na2SO4 0.05 g 0.05 0.352
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) and dispense medium under air atmosphere into culture vessels suitable for anaerobic cultivation (e.g., 20 ml medium per 100 ml serum bottle), seal with grey butyl rubber stoppers and autoclave. Add HEPES buffer and ammonium chloride from stock solutions sterilized by autoclaving. Trace elements, pyruvate, bicarbonate, and vitamins are added from sterile stock solutions sterilized by filtration. Adjust pH of the complete medium to 7.5 using a sterile solution of bicarbonate (5% w/v) or 1 N HCl.
1630 NITROSOSPHAERA MEDIUM