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Main sol. 946a

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Internal Solution-ID: S1958

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Synthetic seawater (2 x conc.) 500.00 ml - -
NH4Cl 0.33 g 0.326 6.102
KH2PO4 0.33 g 0.326 2.399
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 1.00 g 0.989 9.332
Yeast extract
(OXOID) 		3.00 g 2.967 -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.60 g 0.593 2.471
Distilled water 500.00 ml - -
1 Dissolve ingredients (except carbonate, yeast extract, vitamins and sulfide), 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. Prior to inoculation add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere and yeast extract, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins are sterilized by filtration. Adjust pH of complete medium to 6.8, if necessary.
946a CALDITHRIX MEDIUM