16S rDNA Sequencing and Microbiota Analysis

Using the forward and reverse primer pairs, sequencing of 16S rDNA was performed. Third-generation microbial diversity was based on the PacBio sequencing platform, and single molecule real-time cell sequencing (SMRT Cell) was used to sequence the marker gene. The circular consensus sequence was filtered, clustered, or denoised, and species annotation and abundance analysis were performed as previously published, revealing the species composition of each sample. The following analyses were performed: analyses of species taxonomy and annotation, analyses of significant differences, and analyses of diversity (alpha and beta diversity).