16s rRNA-Seq of mouse: P8_5
. The genomic DNA in mouse stool samples was extracted using the E.Z.N.A. Stool DNA Kit (D4015, Omega, Inc., USA) according to manufacturers instructions. Then, the total DNA being extracted was added into the 50 L of Elution buffer for dilution. The hypervariable region of the 16S rRNA V4V5 was amplified using forward 5'-GTGCCAGCMGCCGCGG-3', and reverse 5'-CCGTCAATTCMTTTRAGTTT-3' primers. PCR products were purified via AMPure XT beads (Beckman Coulter Genomics, Danvers, MA, USA), and quantified by Qubit (Invitrogen, USA). The size of amplicon library was assessed by Agilent 2100 Bioanalyzer (Agilent, USA), and the quantity was evaluated by Library Quantification Kit for Illumina (Kapa Biosciences, Woburn, MA, USA).