Sequence reads of fecal microbiome from SS, SR and Con C57BL/6J mice

The composition of the fecal microbiome was analyzed using high-throughput MiSeq sequencing. Sequencing experiments involved the following steps: DNA extraction, PCR amplification and product purification, real-time quantitative PCR, MiSeq library construction, and MiSeq sequencing. PCR amplification was carried out using TransStart FastPfu DNA Polymerase (TransGen Biotech, Beijing, China) and an ABI GeneAmp 9700 system (Thermo Fisher Scientific, Wilmington, DE, USA). Bacterial DNA fragments were amplified using forward primer-338 and reverse primer-806. The DNA was quantified using a Nanodrop spectrophotometer (Thermo Fisher Scientific), and paired-end sequencing of the V3-V4 region of 16S rRNA was performed using an Illumina MiSeq system.

本研究采用高通量MiSeq测序技术，对粪便微生物组的组成进行了分析。本次测序实验包含如下流程：DNA提取、PCR扩增与产物纯化、实时定量PCR、MiSeq文库构建以及MiSeq测序。PCR扩增采用TransStart FastPfu DNA聚合酶（TransStart FastPfu DNA Polymerase，北京全式金生物技术有限公司，TransGen Biotech，中国北京）以及ABI GeneAmp 9700型扩增系统（赛默飞世尔科技，Thermo Fisher Scientific，美国特拉华州威尔明顿）。采用正向引物338与反向引物806扩增细菌DNA片段。本研究使用Nanodrop分光光度计（Nanodrop spectrophotometer，赛默飞世尔科技，Thermo Fisher Scientific）对DNA进行定量，并通过Illumina MiSeq平台完成16S rRNA的V3-V4可变区双端测序。