Cecal Microbiome Profiling via 16S rDNA Sequencing

Total genomic DNA samples of cecal digesta were extracted using a commercial kit (51306, QIAGEN, Hamburg, Germany). The PCR amplification of the V3–V4 region of the 16S rDNA gene of the cecal microbiota was performed using the forward primer 338F (5'-ACTCCTACGGGAGGCAGCA-3') and the reverse primer 806R (5'-GGACTACHVGGGTWTCTAAT-3'), and the 16S rDNA gene amplicon sequencing was performed using the Illumina TruSeq Nano DNA LT Library Prep at Suzhou PANOMIX Biomedical Tech Co., Ltd. (Suzhou, China). The bioinformatics analysis of the sequencing data was performed using QIIME2 (2019.4) and R packages (v3.2.0) with slight modifications based on the official tutorials (https://docs.qiime2.org/2019.4/tutorials/). The detailed sequencing and analysis methods were presented in our previous study [17 (link)].