As already described in Gazzola et al. (2022) (link), anaerobic digestate samples were collected at different sampling times to perform the analysis of microbial community composition over the reactor operation. A small aliquot (2 mL) was centrifuged at 15,000 rpm for 2 min, the resulting pellet was immediately stored at −20°C and used for DNA extraction with DNeasy PowerSoil Pro Kit (QIAGEN - Germantown, MD) following the manufacturer’s instructions. After checked the quality (1.6 < A260/280 < 1.8 and A260/230 > 2) with a Nanodrop 3,300 (Thermo Scientific, Italy), the genomic DNA was used for the high-throughput sequencing of 16S rRNA gene. The V1-V3 region of bacterial 16S rRNA gene (27F 5′- AGAGTTTGATCCTGGCTCAG-3; 534R 5-ATTACCGCGGCTGCTGG-3) was sequenced following library preparation and protocol described in Crognale et al. (2019) (link). The samples were paired end sequenced (2 × 301bp) on a MiSeq platform (Illumina) using a MiSeq Reagent kit v3, 600 cycles (Illumina, United States) following the standard guidelines for preparing and loading samples, with 20% of Phix control library. Bioinformatic analysis were performed using QIIME2 v. 2018.2 (Bolyen et al., 2019 (link)) as described in Crognale et al. (2021) (link). The Dataset is available through the Sequence Read Archive (SRA) under accession PRJNA1052454.
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