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).