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Main sol. 1171

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Internal Solution-ID: S2343

Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 2.000 g 2 11.483
NH4Cl 0.500 g 0.5 9.348
NaCl 120.000 g 120 2053.39
MgSO4 x 7 H2O 0.500 g 0.5 2.029
NaS2O3 x 5 H2O 0.024 g 0.024 -
Trace element solution SL-6 1.000 ml - -
NaHCO3 1.700 g 1.7 20.236
Vitamin B12 100.000 µg 1.0e-4 7.4e-5
Methanol 2.000 ml 1.584 49.435
Distilled water 1000.000 ml - -
1 pH 7.5
2 Prepare the medium without methanol, vitamin B12 MgSO4 x 7 H2O, NaHCO3 and NaS2O3 x 5 H2O in closed vessels with a medium to headspace ratio of 1:5 to 1:10. Add the methanol, MgSO4 x 7 H2O, NaHCO3 and NaS2O3 x 5 H2O from autoclaved stock solutions after sterilisation of the medium. Vitamin B12 is added from a filter sterilised stock solution to the sterile medium. Incubate the strains with shaking at 150 rpm. Growth is faster in the presence of 20mM sodium acetate instead of methanol.
1171 METHYLOHALOMONAS MEDIUM