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Main sol. 731

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Internal Solution-ID: S1485

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.30 g 0.297 1.704
KCl 0.30 g 0.297 3.98
NaCl 0.50 g 0.495 8.463
NH4Cl 1.00 g 0.989 18.492
MgCl2 x 6 H2O 0.30 g 0.297 1.46
CaCl2 x 2 H2O 0.05 g 0.049 0.336
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract 10.00 g 9.891 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 3.00 g 2.967 35.323
Na2CO3 3.00 g 2.967 27.997
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.15 g 0.148 0.845
Na2S x 9 H2O 0.50 g 0.495 2.059
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, carbonate, vitamins, cysteine and sulfide. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dissolve carbonate and bicarbonate and adjust pH of medium to 8.5 using HCl. Dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration.
731 THERMOSYNTROPHA MEDIUM