16S rDNA amplicon sequencing of human fecal sample NM24D0905
DNA was extracted from human fecal samples using the QIAsymphony DSP Virus/Pathogen Midi Kit (Qiagen, catalog no. 937055). The V2, V3, V4, V6-V7, V8, and V9 hypervariable regions of bacterial 16S rDNA were amplified by PCR using the Ion 16S Metagenomics Kit (Thermo Fisher Scientific, catalog no. A26216) and Phusion Hot Start Flex 2X Master Mix. Amplicon libraries were adapter-ligated, amplified by emulsion PCR, and sequenced as single-end reads on an Ion Torrent PGM. Read lengths were variable up to approximately 400 bp depending on amplicon length. FASTQ files were converted from final BAM files and contain quality-filtered reads after adapter and primer trimming and removal of low-quality reads.