PB-18-1
DNA was extracted using the QiaAMP Power Fecal Pro kit (Qiagen) according to the manufacturers instructions. DNA concentration was normalized and 16S rRNA amplicon libraries were prepared using primers to amplify the V3-V4 region of the bacterial 16S rRNA gene, with Illumina adapters incorporated as described in the Illumina 16S rRNA Metagenomic Library Preparation guide, with the exception that 30 amplification cycles were used. Following index PCR and purification, the products were quantified using the Qubit high sensitivity DNA kit (Life Technologies) and pooled equimolarly. The pooled libraries were assessed using an Agilent high sensitivity DNA kit and examined by quantitative PCR (qPCR) using the Kapa Quantification kit for Illumina (Kapa Biosystems, USA) according to the manufacturers guidelines. Libraries were then diluted and denatured following Illumina guidelines and sequenced (2300bp) on the Illumina MiSeq platform.