Microbiome analysis in mice fed HFD or HFD-Soybean protein isolate. mouse gut metagenome

Male C57BL/6 mice were randomly assigned to high-fat diet (HFD) or soybean protein isolate-containing HFD (HFD-SPI) for 4 weeks in a factorial design. Fecal droppings were collected at the end of treatment. Fecal DNA was extracted from frozen samples using the FastDNA SPIN Kit for Feces (MP Biomedicals, Santa Ana, CA) per the manufacture's instructions.The V4 region of the 16S rRNA gene was amplified using dual-indexed 515F/806R primers.he amplicons were sequenced using an Illumina MiSeq with a MiSeq Reagent 222 kit V3 (Illumina, USA).The libraries were prepared following the Illumina protocol for 10pM: 'Preparing Libraries for Sequencing on the MiSeq' (part 15039740, Rev. D).