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Main sol. 1718

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Internal Solution-ID: S3555

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.20 g 0.199 1.465
NH4Cl 0.25 g 0.249 4.66
NaCl 1.00 g 0.997 17.06
MgSO4 x 7 H2O 0.40 g 0.399 1.618
KCl 0.50 g 0.499 6.687
CaCl2 x 2 H2O 0.15 g 0.15 1.017
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Na-acetate 0.80 g 0.798 9.723
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.997 9.407
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, vitamins and sulfide) and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of the complete medium to 6.8 - 7.0, if necessary.
2 After inoculation, pressurize culture vessels with sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
1718 METHANOSPIRILLUM PURGAMENTI MEDIUM