16s-seq of mouse
DNA Extraction and 16S rRNA Sequencing; total DNA was extracted using PowerSoil pro DNA Extraction kit (Qiagen, Hilden, Germany), following the manufacturer's instructions. This included mechanical lysis steps by bead-beating and enzymatic digestion to ensure effective disruption of bacterial cell walls and maximize DNA yield. DNA quality and concentration were assessed using a plat reader and fluorometrically with a Qubit fluorometer (Thermo Fisher Scientific, MA, USA). A sequencing library was then prepared targeting the hypervariable V3V4 regions of the 16S rRNA according to Illuminas amplicon library preparation protocol for the MiSeq System using primer pair-F(5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG-3),R(5GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC-3)