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Main sol. 374

Internal Solution-ID: S714

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.499 3.663
MgSO4 x 7 H2O 0.40 g 0.399 1.618
NaCl 0.40 g 0.399 6.824
NH4Cl 0.25 g 0.249 4.66
CaCl2 x 2 H2O 0.05 g 0.05 0.339
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Na-acetate 1.00 g 0.997 12.154
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.997 9.407
Isopropyl alcohol
(25% v/v)
30.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 970.00 ml - -
1 Dissolve ingredients except carbonate, isopropyl alcohol and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add isopropyl alcohol and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 7.0 - 7.2, if necessary.
374 METHANOSPIRILLUM SK MEDIUM