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Main sol. 846a

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Internal Solution-ID: S1734

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.30 g 0.299 5.592
K2HPO4 0.20 g 0.199 1.145
KH2PO4 0.30 g 0.299 2.198
CaCl2 x 2 H2O 0.15 g 0.15 1.017
KCl 0.50 g 0.499 6.687
NaCl 20.00 g 19.94 341.208
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 2.00 g 1.994 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
MgCl2 x 6 H2O 3.00 g 2.991 14.712
Na2CO3 1.50 g 1.496 14.11
L-Serine 1.05 g 1.047 9.962
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.499 2.838
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except magnesium chloride, carbonate, serine, vitamins, cysteine, and sulfide), 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 to 30% of their volume and autoclave. Add magnesium chloride, serine, vitamins, cysteine, 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 atmosphere. The vitamin solution should be sterilized by filtration. Adjust the pH of the complete medium to 7.2, if necessary.
846a AMINOBACTERIUM MEDIUM