Gut Microbiome Analysis of Mice

Feces samples derived from all mice were collected using a Qiagen stool kit at nine o’clock in the morning and immediately transferred to a −80 °C freezer. The total DNA was extracted using a previously published method [19 (link)]. The V4-V5 region of the bacterial 16S rRNA gene was amplified using specific PCR primers (515F 5′-GTGCCAGCMGCCGCGGTAA-3′, R926 5′-CCGTCAATTCMTTTRAGT-3′). The sequencing was performed on the Ion PGM™ platform [20 (link)] according to the protocols of the BGI-Shenzhen laboratory. The original 16S rRNA sequencing reads were deposited at the European Bioinformatics Institute (EBI) databases under the accession ID ERP113248.