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.
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