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Main sol. 611

Internal Solution-ID: S1311

Equipment needed: Autoclave Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
Synthetic seawater mix 125.00 ml - -
Modified Wolin's mineral solution 10.00 ml - -
Fe(NH4)2(SO4)2 x 6 H2O
(0.01% w/v)
2.00 ml 2.0e-4 -
NH4Cl 0.25 g 0.25 4.674
KH2PO4 0.07 g 0.07 0.514
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
KNO3 1.00 g 1 9.891
Yeast extract 0.50 g 0.5 -
NaHCO3 2.00 g 2 23.808
Distilled water 875.00 ml - -
1 Dissolve ingredients (except nitrate, yeast extract and bicarbonate) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Adjust pH to 7.0 and dispense under 100% N2 gas atmosphere into serum vials (e.g., 20 ml medium per 100 ml bottle). Replace gas phase with 80% H2 and 20% CO2 gas mixture and autoclave. Prior to inoculation add nitrate and yeast extract from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.0 - 7.2.
611 PYROBACULUM (BS) MEDIUM