16s rRNA sequencing of NARMS Poultry Cecum Sample
Total non-enriched extracted DNA from each sample was subjected to 16S rRNA gene copy number quantification using qPCR prior to library preparation. For 16S sequencing, the target copy number threshold was set to at least 167,000 molecules / l. Amplification of libraries was performed using a dual-indexing Illumina primer set (Forward primer:  5TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGAGGCAGCAG3 and Reverse primer: 5GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGGACTACHVGGGTWTCTAAT 3) targeting the V3-V4 region (Gohl et al., 2016). Amplicons were quantified using a PicoGreen dsDNA assay kit (Life Technologies, Carlsbad, CA, USA). Sequencing was performed at the University of Minnesota Genomics Center (UMGC) using Illuminas V3 chemistry (2x300 bp paired-end reads) on the Illumina MiSeq platform (San Diego, CA, USA). All libraries were sequenced on the same sequencing run, which generated a mean 76.7k reads per sample.
SUB16024763
210619483