Shotgun metagenome sequencing of QIH-induced mice

This experiment was performed to investigate the effects of QIH on the cecal and small intestinal microbiota. The intestinal microbiota were compared in Qrfp-iCre mice at multiple time points: before QIH induction (0 h), during the QIH state (18 h and 36 h), and after recovery (168 h). Genomic DNA was extracted from intestinal contents using the QIAamp PowerFecal Pro DNA Kit (Qiagen N.V., Hilden, Germany). The extracted DNA was used to prepare sequencing libraries with the NEBNext Ultra II FS DNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) and NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1 and Set 2) following the manufacturer's instructions. Library quality was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA). Libraries were sequenced on a HiSeq system with a 2 Ã? 150 bp paired-end protocol (Azenta Life Sciences, Burlington, MA).

本实验旨在探究QIH对盲肠与小肠菌群的影响。本研究以Qrfp-iCre小鼠为实验对象，在多个时间点对其肠道菌群进行比较分析：QIH诱导前（0 h）、QIH状态期间（18 h与36 h）以及恢复阶段（168 h）。采用德国希尔德市Qiagen N.V.公司的QIAamp PowerFecal Pro DNA试剂盒，从肠道内容物中提取基因组DNA。参照制造商说明书，使用适配Illumina平台的NEBNext Ultra II FS DNA Library Prep Kit试剂盒与NEBNext Multiplex Oligos for Illumina（双索引引物Set 1与Set 2）对提取得到的DNA进行测序文库构建。采用Agilent 2100 Bioanalyzer生物分析仪（美国加利福尼亚州圣克拉拉市Agilent Technologies, Inc.公司）对文库质量进行检测评估。随后采用HiSeq测序系统，以2×150 bp双端测序方案完成测序，测序服务由美国马萨诸塞州伯灵顿市Azenta Life Sciences提供。