Microbiome analysis in mice fed HFD or HFD-Soybean protein isolate

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).

将雄性C57BL/6小鼠采用析因设计随机分为两组，分别饲喂高脂膳食（high-fat diet, HFD）及添加大豆分离蛋白（soybean protein isolate, SPI）的高脂膳食（HFD-SPI），造模周期为4周。造模结束后收集粪便样本。使用FastDNA SPIN Kit for Feces粪便基因组提取试剂盒（MP Biomedicals，美国加利福尼亚州圣安娜市），严格遵循制造商说明书，从冻存粪便样本中提取粪便DNA。采用双索引引物515F/806R扩增16S rRNA基因的V4区。扩增子使用Illumina MiSeq测序平台及MiSeq Reagent 222 Kit V3测序试剂盒（美国Illumina公司）进行测序。文库构建严格按照Illumina官方发布的10pM浓度文库制备方案《MiSeq测序文库制备指南》（文档编号15039740，修订版D）执行。