SCFAs supplementation exerts metabolic benefits. mouse gut metagenome

The 7-week-old C57BL/6J mice were fed on a HFD diet or HFD containing 5% cellulose, acetate, propionate, butyrate for 4 weeks in a factorial design. Feces were collected at after treatments. Fecal DNA was extracted from frozen samples using the FastDNA SPIN Kit for Feces by following the manufacturer's instructions. Partial 16S rRNA gene sequences were amplified targeting the hypervariable regions V4 using 515F/806R primers. The amplicons were sequenced using an Illumina MiSeq with MiSeq Reagent 222 kit V3. The libraries were prepared following the Illumina protocol for 8pM: 'Preparing Libraries for Sequencing on the MiSeq'.

本实验采用析因设计，将7周龄C57BL/6J小鼠分为两组，分别饲喂高脂饮食（High Fat Diet, HFD）以及添加5%纤维素、乙酸盐、丙酸盐、丁酸盐的高脂饮食，持续干预4周。处理结束后收集粪便样本。采用FastDNA SPIN Kit for Feces试剂盒，按照制造商说明书从冷冻粪便样本中提取粪便DNA。以515F/806R为引物，靶向扩增16S核糖体RNA（16S rRNA）基因的V4高变区片段，获得扩增产物。使用搭载MiSeq Reagent 222 Kit V3试剂的Illumina MiSeq测序平台对扩增产物进行测序。文库构建严格遵循Illumina针对8pM浓度的建库方案：《Preparing Libraries for Sequencing on the MiSeq》。