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Main sol. 663

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Internal Solution-ID: S1395

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 5.44 g 5.402 39.697
MgCl2 x 2 H2O 0.18 g 0.179 1.362
CaCl2 x 2 H2O 0.03 g 0.03 0.203
NH4Cl 0.50 g 0.497 9.283
Trace element solution 5.00 ml - -
Yeast extract 1.00 g 0.993 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na-pyruvate 2.50 g 2.483 22.56
Na2S2O3 x 5 H2O 2.00 g 1.986 8.002
Wolin's vitamin solution (10x) 2.00 ml - -
L-Cysteine HCl x H2O 0.40 g 0.397 2.262
Na2S x 9 H2O 0.40 g 0.397 1.654
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except pyruvate, thiosulfate, vitamins, cysteine and sulfide), then sparge medium with 100% N2 gas 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 pyruvate, thiosulfate, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of pyruvate and vitamins are sterilized by filtration. Adjust medium pH to 7.5 by adding sterile anoxic 1 N NaOH.
663 DESULFITOBACTERIUM MEDIUM