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Main sol. 981

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Internal Solution-ID: S2010

Equipment needed: Filter Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 19.00 g 18.943 324.147
MgCl2 x 6 H2O 9.00 g 8.973 44.137
MgSO4 x 7 H2O 0.15 g 0.15 0.607
CaCl2 x 2 H2O 0.30 g 0.299 2.035
KCl 0.50 g 0.499 6.687
KH2PO4 0.42 g 0.419 3.077
(NH4)2SO4 0.10 g 0.1 0.755
NaBr 0.05 g 0.05 0.485
SrCl2 x 6 H2O 0.02 g 0.02 0.075
KNO3 1.01 g 1.007 9.96
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Na2CO3 1.00 g 0.997 9.407
Yeast extract 0.10 g 0.1 -
DL-Malic acid 1.34 g 1.336 9.963
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.05 g 0.05 0.284
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, yeast extract, malic acid, vitamins 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. Prior to inoculation add yeast extract, malic acid, vitamins (sterilized by filtration), and cysteine 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 6.2.
981 GEOTHERMOBACTER MEDIUM