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Main sol. 778b

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Internal Solution-ID: S1583

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.33 g 0.329 2.418
NH4Cl 0.33 g 0.329 6.151
KCl 0.33 g 0.329 4.413
MgCl2 x 6 H2O 0.33 g 0.329 1.618
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
NiCl2 x 6 H2O
(0.1% w/v) 		0.20 ml 2.0e-4 0.002
Yeast extract 1.00 g 0.997 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 2.50 g 2.493 29.67
CaCl2 x 2 H2O 0.33 g 0.329 2.238
Na2-fumarate 1.60 g 1.595 9.968
Na2S2O3 x 5 H2O 2.00 g 1.994 8.034
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 bicarbonate, calcium chloride, fumarate, thiosulfate, vitamins and sulfide). Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, add solid bicarbonate and adjust pH to 7.2. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add calcium chloride, fumarate, thiosulfate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Stock solutions of fumarate, thiosulfate and vitamins are sterilized by filtration. Adjust pH of complete medium to 7.0 - 7.2, if necessary.
778b CALDIMICROBIUM MEDIUM