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展开分析。测序实验涵盖以下步骤：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（全式金生物技术有限公司，中国北京）与ABI GeneAmp 9700扩增系统（赛默飞世尔科技，美国特拉华州威尔明顿）。研究使用正向引物338与反向引物806扩增细菌DNA片段，通过Nanodrop分光光度计（赛默飞世尔科技）对DNA进行定量，并依托Illumina MiSeq系统完成16S rRNA V3-V4可变区的双端测序。