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Main sol. 903

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Internal Solution-ID: S1856

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.499 3.663
MgCl2 x 6 H2O 0.33 g 0.329 1.618
NaCl 0.40 g 0.399 6.824
CaCl2 x 2 H2O 0.05 g 0.05 0.339
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Trypticase peptone
(BD BBL) 		0.25 g 0.249 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.496 14.11
Sucrose 5.00 g 4.985 14.563
Seven vitamins solution 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.703
Na2S x 9 H2O 0.30 g 0.299 1.245
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, sucrose, vitamins and reducing agents), then 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. Add sucrose, cysteine 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 are prepared under 100% N2 gas and sterilized by filtration. The pH of the complete medium should be at 7.0.
903 CALDANAEROBIUS MEDIUM