Microbial community CSTR Cellulose Reaactor 2
Digestate samples (5 ml) were collected at the beginning of the experiment (inoculum, Day 0) and at the end of the AD process (Day 52) from each reactor. To investigate functional dynamics of microbial communities, extraction was performed immediately after collection to minimise RNA degradation and preserve the integrity. RNA extraction was performed using the RNeasy PowerFecal Pro kit (Qiagen, Germany) according to manufacturers guidance. The purity (A260/A280 and A260/A230) of the extracted RNA was quantified using a Nanodrop 1000 spectrophotometer (Thermo Fisher Scientific), and the concentration of RNA was measured using a Qubit RNA BR assay kit (Thermo Fisher Scientific). The absence of DNA in the RNA extracts was verified by 1 % (w/v) 1 TAE agarose gels. Extractions for each reactor were carried out in duplicate; replicates were then pulled together to minimise extraction variability. RNA samples were paired-end sequenced (2  150bp) on the Illumina NovaSeq (PE150) platform at Novogene Co., Ltd (Cambridge, UK).