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Main sol. 1304

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Internal Solution-ID: S2606

Equipment needed: Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
MgCl2 x 6 H2O 0.2 g 0.199 0.981
CaCl2 x 2 H2O 0.1 g 0.1 0.678
NaNO3 1.7 g 1.695 19.941
(NH4)2SO4 0.1 g 0.1 0.755
KH2PO4 0.1 g 0.1 0.733
KCl 0.1 g 0.1 1.337
Trace element solution SL-10 1.0 ml - -
Selenite-tungstate solution 1.0 ml - -
Na2S2O3 x 5 H2O 0.4 g 0.399 1.607
NaHCO3 5.0 g 4.985 59.341
Wolin's vitamin solution 1.0 ml - -
Distilled water 1000.0 ml - -
1 Dissolve ingredients except trace elements, thiosulfate, bicarbonate and vitamins, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to remove dissolved oxygen and to saturate the solution with CO2. Dispense solution under 80% N2 and 20% CO2 gas atmosphere in vials suitable for anaerobic cultures (e.g., Balch-type tubes) to 30% of volume. Close vials with butyl rubber septa and autoclave. Add trace element solution, selenite-tungstate solution and vitamins 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. The vitamin solution should be sterilized by filtration. Adjust pH of the complete medium to 6.7 - 7.0, if necessary.
2 After inoculation, add 1 bar overpressure of sterile 100% H2 gas.
3 Note: Use at least 10% (v/v) as inoculum.
1304 SULFURITALEA MEDIUM