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Main sol. 778

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Internal Solution-ID: S1581

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.33 g 0.328 2.41
NH4Cl 0.33 g 0.328 6.133
KCl 0.33 g 0.328 4.4
MgCl2 x 6 H2O 0.33 g 0.328 1.614
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
NiCl2 x 6 H2O
(0.1% w/v) 		0.20 ml 2.0e-4 0.002
Yeast extract 1.00 g 0.994 -
Na2-9,10-anthraquinone-2,6-disulfonate
(TCI A0308) 		8.25 g 8.201 -
NaHCO3 2.50 g 2.485 29.582
CaCl2 x 2 H2O 0.33 g 0.328 2.231
Glycerol 2.60 ml - -
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, calcium chloride, glycerol and vitamins), boil medium for some minutes to dissolve the anthraquinone, then cool under 80% N2 and 20% CO2 gas atmosphere to room temperature. Add solid bicarbonate and adjust pH to 6.8, dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add calcium chloride, glycerol and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Vitamins are sterilized by filtration. Adjust pH of complete medium to 6.8, if necessary.
778 THERMOTERRABACTERIUM MEDIUM