title: RNA-Seq of mice feces: adult male feces
design: 200mg of fresh feces were collected 1 month after operation (11 months of age). Intestinal microbial DNA was extracted with a DNA kit (Solarbio, China), and the sequence of the V3-V4 region was amplified by PCR (Forward primer CCTACGGGNGGCWGCAG, Reverse primer GACTACHVGGGTATCTAATCC). PCR products were recovered by AGAR gel electrophoresis, then purified and quantified. High-throughput sequencing was performed on the Illumina NovaSeq 6000. QIIME2.0 software package conducted OTU clustering for the base sequence fragments in the V3-V4 region. After comparison, the corresponding taxon of OTU was obtained, and various correlation analysis was performed based on OTU.