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Incubate at 42°C without shaking. Activity and growth can be followed by measuring the depletion of ammonium or the production of nitrite and/or nitrate. Higher ammonium concentrations are inhibitive. For that reason our recommendation is follow nitrite content. As soon as nitrite has been produced and is depleting again (to form nitrate) another dose of ammonium can be added. Incubation time may be up to several weeks. We checked the nitrite content (absence/presence) every 5-7 days and added ammonium chloride about every 2-3 weeks. Don't expose cultures to intense light.
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