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Main sol. 1277

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Internal Solution-ID: S2555

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 0.25 g 0.249 4.265
MgCl2 x 6 H2O 0.20 g 0.199 0.981
CaCl2 x 2 H2O 0.10 g 0.1 0.678
NaNO3 1.70 g 1.695 19.941
(NH4)2SO4 0.10 g 0.1 0.755
KH2PO4 0.10 g 0.1 0.733
KCl 0.10 g 0.1 1.337
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Na2CO3 1.50 g 1.496 14.11
Na2S2O3 x 5 H2O 2.50 g 2.493 10.043
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, thiosulfate and vitamins) and 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 to 30% volume and autoclave. Add thiosulfate and vitamins 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. Stock solutions of thiosulfate and vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.5, if necessary.
2 After inoculation add sterile air in an amount equivalent to 1% (v/v) of the total volume of the culture vessel.
1277 SULFURICELLA MEDIUM