Gut Microbiome Analysis of Mice

The colon contents were collected from all mice immediately after sacrifice. The V3–V4 region of the 16sRNA gene was sequenced in the 48 content samples. Microbial DNA was extracted from the colon contents using the QIAamp DNA Stool Mini Kit (Qiagen, Hilden, Germany). The microbial V3-V4 region was amplified by PCR using the primers 5′-ACTCCTACGGGAGGCAGCA and 3′-GGACT ACHVGGGTWTCTAAT. Amplicons were extracted from a 2% agarose gel using the TIANgel MIdi Purification Kit (TIANGEN BIOTECH, Beijing, China) and were subsequently purified. For detailed experimental steps on Illumina MiSeq sequencing, please see our previous article (Zhu et al., 2018 (link)); The general data analysis was performed by a commercial company (Novogene, Beijing, China).