Total genome DNA of gut samples was extracted using the kit (Tiangen Company, Beijing, China). The DNA concentration was performed using Qubit@ 2.0 Fluorometer (Thermo Scientific). DNA quality was also monitored on 1% agarose gels. PCR was performed to produce V4 regions of the 16S rDNA gene using the conserved primers 515F (5'-GTGCCAGCMGCCGCGGTAA-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'). PCR products and operate electrophoresis on 2% agarose gel for detection. Samples with bright main strip between 400-450bp were chosen for purifing with GeneJET Gel Extraction Kit (Thermo Scientific). PCR fragments were sequenced by Novogene Bioinformatics Technology Co., Ltd. (Tianjin, China). Libraries was sequenced on an Illumina HiSeq platform and 250 bp paired-end reads were generated.
GEO Accession
GSM3263523