16S rRNA seq of mus musculus: cecal feces & WT-DSS2
Total genome DNA were extracted from cecal contents, and then the purity and concentration of all DNA samples were tested. According to the selection of sequencing region, the selected V3-V4 variable region was amplified by PCR using specific primers with Barcode and high-fidelity DNA polymerase. PCR products were recovered and quantified with the Microplate reader (BioTek, FLx800) fluorescence quantitative system, and the corresponding proportions were mixed according to the sequencing requirements of each sample. Sequencing was conducted on the Illumina MiSeq platform (Illumina, San Diego, CA, USA). The Quantitative Insights into Microbial Ecology (QIIME2, 2020.11) pipeline was used to analyze the generated and demultiplexed sequences. All representative reads were annotated and blasted against Silva database Version 138 (or Unite) (16s/18s/ITS rDNA) using q2-feature-classifier with the default parameters