Microbial diversity of anaerobic digesters at different temperature regimes
Pellet (200 mg) of each sample was collected for DNA extraction in duplicate with the FastDNA SPIN Kit for Soil. For the amplification of bacterial 16S rRNA gene, the 16S universal Eubacterial primers 27F (AGRGTTTGATCMTGGCTCAG) and 519R (GTNTTACNGCGGCKGCTG) with 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 and purified using Agencourt Ampure beads. Purified and pooled samples were loaded to the Roche 454 FLX titanium instrument.