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Main sol. 1210b

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Internal Solution-ID: S2413

Equipment needed: Autoclave Hungate tubes Centrifuge Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 18.00 g 17.946 307.087
MgCl2 x 6 H2O 4.00 g 3.988 19.616
KCl 0.33 g 0.329 4.413
CaCl2 x 2 H2O 0.33 g 0.329 2.238
(NH4)2SO4 0.50 g 0.499 3.773
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1 % w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.997 9.407
KH2PO4 0.33 g 0.329 2.418
FeCl2 x 4 H2O 0.52 g 0.518 2.608
Yeast extract 0.20 g 0.199 -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.10 g 0.1 0.568
Amorphous Fe(OH)3 100.00 ml - -
Distilled water 900.00 ml - -
1 Dissolve ingredients except carbonate, hydrogenphosphate, ferrous chloride, yeast extract, vitamins, cysteine and ferric iron hydroxide sludge. Suspend pellet of ferric iron hydroxide in medium and sparge with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Thereafter, dispense suspension under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave for 30 min. Add hydrogenphosphate, ferrous chloride (dissolved in 0.1 N HCl), yeast extract, cysteine and vitamins 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. Vitamins and ferrous chloride should be sterilized by filtration. The pH of the complete medium should be at 5.5.
1210b PYRODICTIUM DELANEYI MEDIUM