16SrRNA-Mus musculus-MFD-Omega3 Diet-Week 24
The 16S rRNA gene amplicon library preparation was performed using Ion 16S Metagenomics Kit (Thermo Fisher Scientific, Carlsbad, CA, USA). The libraries were generated from 20 ng of fecal DNA, using a combination of two pools of primers targeting the V2, V4 and V8 hypervariable regions of 16S rRNA gene with the pool 1, and the V3, V6-7 and V9 regions with the pool 2. Primers were partially digested and bar-codes were ligated to the amplicons, purified using the Agencourt AMPure XP beads (Beckman Coulter; Pasadena, CA, USA) according to the manufacturers protocol, and stored at  20 C. The concentration of each 16S library was determined by qPCR using the Ion Universal Library Quantitation Kit (Thermo Fisher Scientific Carlsbad, CA, USA). The library was diluted to ~ 10 pM before template preparation. Template preparation of the barcoded libraries was performed using the Ion PGM Hi-Q View OT2 kit (Thermo Fisher Scientific Carlsbad, CA, USA) and the Ion OneTouch 2 System (Thermo Fisher Scientific Carlsbad, CA, USA). A maximum of 12 barcoded 16S samples was sequenced on an Ion 316v2 chip using the Ion PGM Hi-Q view Sequencing Kit (Thermo Fisher Scientific; Carlsbad, CA, USA) according to the manufacturers instructions on the Ion Torrent Personal Genome Machine (PGM) platform (Thermo Fisher Scientific Carlsbad, CA,USA).