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Main sol. 899

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Internal Solution-ID: S1851

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.99 18.51
K2HPO4 0.30 g 0.297 1.705
KH2PO4 0.30 g 0.297 2.183
MgCl2 x 6 H2O 1.00 g 0.99 4.87
CaCl2 x 2 H2O 0.10 g 0.099 0.673
KCl 0.20 g 0.198 2.656
NaCl 0.60 g 0.594 10.165
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract 2.00 g 1.98 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.99 9.342
D-Glucose 4.00 g 3.96 21.983
Na2S2O3 x 5 H2O 2.50 g 2.475 9.973
L-Cysteine HCl x H2O 0.50 g 0.495 2.819
Na2S x 9 H2O 0.45 g 0.446 1.855
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, glucose, thiosulfate, 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 same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose, thiosulfate (sterilized by filtration), 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 mixture. The pH of the complete medium should be at 7.0.
899 CALDANAEROBACTER MEDIUM