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Main sol. 1329

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Internal Solution-ID: S2664

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
MgCl2 x 6 H2O 3.05 g 3.041 14.957
CaCl2 x 2 H2O 0.15 g 0.15 1.017
NH4Cl 0.54 g 0.538 10.065
NaCl 20.00 g 19.94 341.208
Na-acetate 6.56 g 6.54 79.728
Trace element solution 2.00 ml - -
Yeast extract
(OXOID) 		1.50 g 1.496 -
Trypticase peptone
(BD BBL) 		0.40 g 0.399 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
KH2PO4 1.19 g 1.186 8.718
K2HPO4 0.21 g 0.209 1.202
Na2CO3 1.50 g 1.496 14.11
2-Mercaptoethanesulfonic acid
(coenzyme M) 		0.14 g 0.14 0.982
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.36 g 0.359 1.494
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except phosphates, carbonate, coenzyme M, vitamins and sulfide) and 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 phosphates, vitamins (sterilized by filtration) 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. Adjust pH of the complete medium to 7.5, if necessary.
1329 METHANOSAETA PELAGICA MEDIUM