110513p2
Raw sequences of filtered water samples: Mississippi River
V4 regions of bacterial 16S rRNA genes were amplified and sequenced using methods described previously (Stone et al. 2016; Payne et al. 2017) that were slightly modified from Kozich et al. 2013. Briefly, DNA was amplified using standard forward (5 -GTGCCAGCMGCCGCGGTAA) and reverse (5 -GGACTACHVGGGTWTCTAAT) primers adapted with dual-index barcodes for Illumina MiSeq next generation sequencing (Kozich et al. 2013), and run through 30 cycles of denaturation (95 C) for 20 s, annealation (55 C) for 15 s, and elongation (72 C) for 2 min, and a final elongation (72 C) for 10 min (Stone et al. 2016; Payne et al. 2017). Amplified DNA was normalized, pooled, and sequenced using an Illumina MiSeq platform located at the Molecular and Genomics Core Facility at the University of Mississippi Medical Center.
110513p2
SUB2197565
110513p2