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Main sol. 311c

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Internal Solution-ID: S579

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.50 g 0.498 9.301
MgSO4 x 7 H2O 0.50 g 0.498 2.019
CaCl2 x 2 H2O 0.25 g 0.249 1.692
NaCl 2.25 g 2.239 38.31
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		2.00 ml 0.002 0.007
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 2.00 g 1.99 -
Casitone 2.00 g 1.99 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
K2HPO4 0.35 g 0.348 1.999
KH2PO4 0.23 g 0.229 1.682
Na2CO3 1.00 g 0.995 9.388
D-Fructose 5.00 g 4.975 27.615
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.7
Na2S x 9 H2O 0.30 g 0.299 1.243
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except phosphates, carbonate, fructose, vitamins, cysteine and sulfide) 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. After autoclaving add phosphates, fructose, vitamins, cysteine and sulfide to the medium 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 fructose and vitamins should be sterilized by filtration. Adjust pH of complete medium to pH 6.5 - 7.0, if necessary.
311c SPOROMUSA ACIDOVORANS MEDIUM