title: Gut microbiome of Aged 21 mice injected with iWAT-EVs without retinoic acid
design: Purified PCR products of fecal bacterial DNA were fragmented, end-repaired, adaptor-ligated, and sequenced on the DNBseq platform using paired-end reads of 150 bp. The PhiX control was added to the sequencing library. Raw reads were first trimmed and quality-checked using TrimGalore v0.5.0. Reads were classified by Kraken2/Bracken to determine bacterial taxonomic compositions. Low abundance OTUs (<0.001%) were removed. The relative abundance at phylum, family, genus, and species levels was analyzed. OTU picking, taxonomic assignment, and construction of phylogenetic trees were followed by downstream statistical analysis to explain differences in community composition between samples or groups via Principal Component Analysis (PCA).