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Main sol. 1250

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Internal Solution-ID: S2482

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 0.45 g 0.449 3.395
MgSO4 x 7 H2O 0.50 g 0.499 2.023
KCl 0.05 g 0.05 0.669
Ca(NO3)2 x 4 H2O 0.02 g 0.02 0.084
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 0.50 g 0.499 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
KH2PO4 10.00 g 9.97 73.264
Na2HPO4 1.00 g 0.997 7.023
D-Fructose 3.50 g 3.49 19.369
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.499 2.838
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except phosphates, vitamins, fructose and cysteine) 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 and autoclave. Prepare a phosphate buffer stock solution with KH2PO4 (25% w/v) and Na2HPO4, adjust pH of the buffer to 5.8. Prior to inoculation add phosphate buffer, vitamins, fructose and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins and D-fructose should be sterilized by filtration. The pH of the complete medium should be at 5.8.
1250 DESULFOSPOROSINUS ACIDOPHILUS MEDIUM