MSCs_Feces_16S_05
Primers were designed targeting conserved regions in 16S rDNA/ITS2 sequences. Total microbial DNA was used for 35 - cycle PCR amplification with Phusion enzyme. PCR products were identified by 1.5% agarose gel electrophoresis, and target fragments were recovered. Purified products were quantified using a Promega QuantiFluor Fluorescence Quantification System with Quant-iT PicoGreen dsDNA Assay Kit and then pooled to construct the library. Paired - end sequencing was carried out on Illumina Sequencing Platform 250PE. Raw 16S sequencing data were normalized by filtering, de - duplication, and chimera filtering in QIIME2 to correct errors. Amplicon Sequence Variants (ASVs) for each microorganism were extracted. The "ggvenn" package was used to create a Venn diagram for ASV visualization, and rarefaction curves were plotted to count ASV abundances.