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Main sol. 1210c

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Internal Solution-ID: S2414

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 18.00 g 17.946 307.087
MgCl2 x 6 H2O 4.00 g 3.988 19.616
KCl 0.33 g 0.329 4.413
CaCl2 x 2 H2O 0.33 g 0.329 2.238
(NH4)2SO4 0.50 g 0.499 3.773
KNO3 1.00 g 0.997 9.861
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
Sulfur
(powder) 		5.00 g 4.985 155.443
Na2CO3 1.00 g 0.997 9.407
KH2PO4 0.33 g 0.329 2.418
Na2S2O3 x 5 H2O 3.50 g 3.49 14.06
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except sulfur, carbonate, hydrogen phosphate, thiosulfate, and vitamins, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur and autoclave at 110°C for 20 min. Add hydrogen phosphate, 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. The pH of the complete medium should be 6.5 - 6.8.
1210c THERMOSULFURIPHILUS MEDIUM