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Main sol. 1255

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Internal Solution-ID: S2489

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 0.988 16.909
NH4Cl 1.00 g 0.988 18.473
MgCl2 x 6 H2O 0.10 g 0.099 0.486
CaCl2 x 2 H2O 0.10 g 0.099 0.672
KH2PO4 0.38 g 0.375 2.759
K2HPO4 0.75 g 0.741 4.255
Trace element solution SL-10 10.00 ml - -
Yeast extract
(OXOID) 		2.00 g 1.976 -
Tryptone
(BD Bacto) 		2.00 g 1.976 -
FeSO4
(0.1% w/v in 0.1 N H2SO4) 		0.50 ml 4.9e-4 0.003
PIPES
(SIGMA) 		3.80 g 3.755 12.418
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
D-Glucose 2.00 g 1.976 10.97
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.494 2.057
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except glucose, vitamins and sulfide), adjust pH to 7.0 and 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. Prior to inoculation add glucose, vitamins and sulfide from sterile anoxic stock solution prepared under 100% N2 gas. Vitamins are sterilized by filtration. The pH of the complete medium should be 7.0.
1255 THERMOVENABULUM MEDIUM