title: 16S rRNA gene amplicon sequencing of layer hen ileal microbiota
design: DNA from ileal luminal content samples was extracted using the Qiagen Powerlyzer soil kit. Samples were then sequenced using 250 bp paired-end reads for each sample of the V4 region of the 16S rRNA gene (515F, 806R; Caporaso et al., 2011; Caporaso et al., 2012) at the Iowa State University DNA Facility using Illumina MiSeq sequencing technology.