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Main sol. 788

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Internal Solution-ID: S1593

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.41 g 0.409 3.004
Na2HPO4 x 2 H2O 0.52 g 0.518 3.652
NH4Cl 2.40 g 2.393 44.734
MgCl2 x 6 H2O 0.40 g 0.399 1.962
Yeast extract 0.20 g 0.199 -
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
CaCl2 x 2 H2O 0.20 g 0.199 1.356
Na2CO3 1.50 g 1.496 14.11
D-Glucose 5.00 g 4.985 27.67
Seven vitamins solution 1.00 ml - -
Na2S x 9 H2O 0.30 g 0.299 1.245
Distilled water 1000.00 ml - -
1 Dissolve ingredients except calcium chloride, carbonate, glucose, vitamins and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add calcium chloride, glucose, vitamins and sulfide 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 should be sterilized by filtration. Prior to use adjust pH of complete medium to 7.2 - 7.4, if necessary.
788 CALORAMATOR MEDIUM