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Main sol. 818

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Internal Solution-ID: S1666

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.20 g 1.196 20.473
KCl 0.30 g 0.299 4.012
NH4Cl 0.30 g 0.299 5.592
KH2PO4 0.20 g 0.199 1.465
Na2SO4 0.30 g 0.299 2.106
MgCl2 x 6 H2O 0.40 g 0.399 1.962
CaCl2 x 2 H2O 0.15 g 0.15 1.017
Trace element solution SL-11 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 0.60 g 0.598 7.121
KNO3 0.50 g 0.499 4.931
Wolin's vitamin solution (10x) 1.00 ml - -
Seven vitamins solution 1.00 ml - -
Na-acetate 0.50 g 0.499 6.077
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, nitrate, vitamins and acetate. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, adjust pH to 7.0 - 7.4, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add nitrate, vitamins and acetate from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins should be sterilized by filtration. The pH of the complete medium should be adjusted to 7.4 - 7.8, if necessary.
818 CHRYSIOGENES MEDIUM