V3-V4 of anaerobic digester inoculum
The extracted DNA was amplified targeting the V3-V4 region of the 16S rRNA gene using primers adapted by Zymo Research (USA) from the standard 341F (CCTACGGGDGGCWGCAG/CCTAYGGGGYGCWGCAG) and 806R (GACTACNVGGGTMTCTAATCC) pair, assessing both Bacteria and Archaea and generating an approximate 460 bp merged fragment. Library preparation was performed with the Quick-16STM Plus NGS Library Prep Kit (V3-V4, Zymo Research), and sequencing was conducted on an Illumina MiSeq platform (100K paired reads).