Shotgun metagenomic sequencing of QIH-induced mice maintained in warm ambient conditions

This experiment aimed to determine whether the characteristic alterations in gut microbiota observed during QIH are dependent on hypothermia. To investigate this, we analyzed the intestinal microbiota of mice in which QIH was induced under warm ambient conditions. The intestinal microbiota were analyzed at multiple time points (0, 18, and 36 h) after QIH induction. Genomic DNA was extracted from intestinal contents using the QIAamp PowerFecal Pro DNA Kit (Qiagen N.V., Hilden, Germany). Sequencing libraries were prepared 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). Sequencing was performed on a HiSeq system using a 2 Ã? 150 bp paired-end protocol (Azenta Life Sciences, Burlington, MA).