title: Viral metagenomics of feces swab samples from laboratory rabbits: rabbitfeces50
design: For viral metagenomic analysis, 40 specimens were collected from 10 laboratory rabbits which were bred in the Animal Center of Jiangsu university (including 10 feces, 10 oral, 10 blood, 10 skin specimens) in October 2018. Before use, swab was put into 1.5 mL centrifuge tubes 500 uL PBS and vortexed for 5 min at room temperature. The supernatants were then collected after centrifugation (5 min, 14,000g). 40 sample supernatants were pooled into 8 pools with 5 sample each according to sample types, where each type of sample included 2 pools. Supernatants were filtered through a 0.45mm 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 (nonviral encapsidated) nucleic acids. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit according to manufacturer's protocol. Eight 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.