NC.M.5.17_F
Fecal microbial DNA was isolated by using QIAamp 96 PowerFecal QIAcube HT Kit (QIAGEN). The 16S V4 region were amplify by the forward primer (F515, 5-GTGCCAGCMG CCGCGGTAA-3) and the reverse primer (R806, 5-GGACTACHVGGGTWTCTAAT-3). The reaction conditions were 3 min at 95 C, followed by 25 cycles of 95 C for 30 s, 55 C for 30 s, 72 C for 30 s, and 5 min at 72 C for a final extension. For library construction, the amplified PCR product were attached with Illumina sequencing adapters by Nextera XT Index Kit then purified by AMPure XP beads. The library quantification was conducted by Bioanalyzer DNA 1000 chip and Agilent Technologies 2100 Bioanalyzer. The sequencing (paired-end reads, 2  494 bp) was performed by Illumina NextSeq.