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Main sol. 1011b

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Internal Solution-ID: S2060

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.33 g 0.329 4.418
MgSO4 x 7 H2O 3.45 g 3.443 13.969
MgCl2 x 6 H2O 2.75 g 2.745 13.5
NaCl 20.00 g 19.96 341.548
CaCl2 x 2 H2O 0.15 g 0.15 1.018
NH4Cl 0.25 g 0.25 4.664
K2HPO4 0.15 g 0.15 0.859
Fe(NH4)2(SO4)2 x 6 H2O 0.01 g 0.01 -
Yeast extract 0.10 g 0.1 -
Trace element solution SL-6 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.998 9.416
D-Glucose 2.00 g 1.996 11.079
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.705
Na2S x 9 H2O 0.30 g 0.299 1.247
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, glucose, 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 and autoclave. Add vitamins, glucose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 6.0 - 6.5, if necessary.
1011b THERMOSIPHO MJ MEDIUM