Rat metagenome Genome sequencing. rat metagenome

For viral metagenomic analysis, 20 wild adult rats were captured from Jiangsu Province, China in 2021, and blood, fecal, and oral pharyngeal swab samples were collected from each rat. Based on the randomization principle, 10 samples were taken from each library. Two libraries of murine blood, two libraries of murine feces, and two libraries of murine oral pharyngeal swabs were obtained. The fecal samples were individually homogenized with a mortar and pestle, resuspended in 1 mL DPBS, and then frozen and thawed rapidly three times on dry-ice. The rest of the samples were freeze-thawed only 3 times repeatedly. The supernatants were then collected after centrifugation (10 min, 15,000 g). Supernatants were filtered through a 0.45-mm filter (Millipore) to remove eukaryotic- and bacterial cell-sized particles, and 200 uL of supernatant from each pool was then subjected to a mixture of nuclease enzymes to reduce the concentration of free (non-viral encapsidated) nucleic acids. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit according to manufacturer's protocol. The six libraries were then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the MiSeq Illumina platform with 250 bases paired ends with dual barcoding for each pool.