feces metagenome Metagenome

Design: For viral metagenomic analysis, fecal samples were collected from 10 black-necked cranes in December 2020 in Sakya County, Shigatse, Tibet. The fecal samples were transported to the laboratory on dry ice and stored at --353.15K before use. Specimens in the specimen pool were homogenized separately using a mortar and pestle, resuspended in 1 mL of DPBS, and then rapidly frozen and thawed three times on dry ice. The supernatant was collected after centrifugation (10 min, 15,000 g). The supernatant was filtered through a 0.45 mm filter (Millipore) to remove eukaryotic and bacterial cell-sized particles, and then 500 liters of supernatant from each pool was treated with a nuclease mixture to reduce the concentration of free (non-viral-encapsulated) nucleic acids. The remaining total nucleic acids were then isolated using the QIAamp MinElute Virus Spin Kit according to the manufacturer's protocol. 1 library was then constructed using the Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced on the Illumina NovaSeq 6000 platform.