Microbial diversity of anaerobic digesters at different temperature regimes
After extraction and purification DNA pellet was fragmented, ligated to sequencing adaptors. For amplification, 16S universal primers Met 86F (GCTCAGTAACACGTGG) and Met 448F (GCGGCGGCTGGCACC) with the barcode sequences, on the forward primers were utilized on single-step 30 cycle PCR, using HotStarTaq Plus Master Mix Kit. Following PCR, amplicon products from different samples were mixed together to prepare library and purified using Agencourt Ampure beads. Pooled library was loaded to a 600 Cycles v3 Reagent cartridge (Illumina) and sequencing was performed on Miseq (Illumina).