Fecal virome of bar-headed geese

For viral metagenomic analysis, fecal samples were collected from 10 bar-headed geese in December 2020 in Sakya County, Shigatse, Tibet. The fecal samples were transported to the laboratory on dry ice and stored at minus 80 degrees before use. Specimens in the specimen pool were homogenized separately using a mortar and pestle, resuspended in 1 milliliter of DPBS, and then rapidly frozen and thawed three times on dry ice. The supernatant was collected after centrifugation. The supernatant was filtered through a 0.45 micron filter (Millipore) to remove eukaryotic and bacterial cell-sized particles, and then 200 microliter of supernatant from each pool was treated with a mixture of nuclease enzymes 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. One library was then constructed using the Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the Illumina NovaSeq 6000 platform with 250 bases paired ends with dual barcoding.