16S rRNA-Seq of mus musculus: adult female cecum and large intestine
According to the instructions of the ISOSPIN Fecal DNA (NIPPON GENE Co., Ltd., Tokyo, Japan), the total genomic DNA from each intestinal content was extracted following the standard protocol. The DNA concentration was quantified using the Qubit 3.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and Qubit dsDNA HS Assay Kit (Life Technologies). The first PCR used the forward primer 340F and the reverse primer 806R. The PCR products were cleaned up, and the amplicon concentration was measured by Qubit system. Second PCR amplification was performed using an index primer of Nextera XT Library Preparation Kit (Illumina Inc., USA). Clean-up and concentration measured again. The PCR products of each sample were mixed to prepare the PCR amplicon libraries. Quality check of each library was conducted by Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA) and Agilent DNA 1000 Kit. After quality assessment and quantification, the same amounts of the amplified products were collected and sequenced by pair-end 2  391 bp in the Illumina Miseq.