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Main sol. 1112

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Internal Solution-ID: S2233

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.54 g 0.538 10.065
KH2PO4 0.14 g 0.14 1.026
MgCl2 x 6 H2O 0.20 g 0.199 0.981
CaCl2 x 2 H2O 0.15 g 0.15 1.017
Na-acetate 0.82 g 0.818 9.966
NaNO3 0.85 g 0.847 9.971
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.997 9.407
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The vitamin solution should be sterilized by filtration. Adjust pH of the complete medium to 7.0, if necessary.
1112 CALDITERRIVIBRIO MEDIUM