qPCR results of extended batch anaerobic digestion at different ammonia concentrations
Ammonia stress to methanogens and syntrophic bacteria in anaerobic digestion could be adaptable and unadaptable, depending on the length of batch anaerobic digestion experiments. Key methanogens and syntrophic bacteria were identified by qPCR in extended batch digestion experiments with active and ammonia-stressed inoculum.
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DNA was then extracted with a DNeasy PowerSoil Kit (Qiagen, Germantown, MD, USA). The extracts were prepared at 20 ng total DNA per reaction mixture and stored at -21°C. Quantitative polymerase chain reaction (qPCR) was performed in 25 µL volumes on a QuantStudio 3 (Life Technologies).