title: Impact of SIV and THC on colonic microbiome changes
design: DNA library was constructed using TruSeq Nano DNA LT Library Preparation Kit (FC-121-4001) and following the manufactures protocol. Briefly, DNA was fragmented by dsDNA Fragmentase (NEB, M0348S) by incubate at 37C for 30min. Library construction begins with fragmented genomic DNA (gDNA). Blunt-end DNA fragments are generated using a combination of fill-in reactions and exonuclease activity, and narrowly size selected with sample purification beads. An A-base is then added to the blunt ends of each strand, preparing them for ligation to the indexed adapters. Each adapter contains a T-base overhang for ligating the adapter to the A-tailed fragmented DNA. These adapters contain the full complement of sequencing primer hybridization sites for single, paired-end, and indexed reads. Single-index adapters are ligated to the fragments and the ligated products are amplified with reduced-bias PCR. Quality control analysis and quantification of the DNA library were performed using agarose gel and KAPA Library Quantification Kits. Paired-end 2x150 sequencing was performed on Illumina NovaSeq sequencing system.