tick metagenome

To investigate the virome of ticks from giant pandas at Dujiangyan Panda Valley, the Chengdu Research Base of Giant Panda Breeding Dujiangyan Field Research Center for Giant Pandas in Sichuan Province, China, our study collected tick specimens between May and July 2020.The samples were collected by disposable materials and resuspended in 1 mL DPBS and vigorously vortexed for 15 min. Before re-suspending in DPBS, samples were homogenized, frozen, and thawed on dry ice three times, the supernatants were then collected after centrifugation. 500 uL of supernatant was filtered through a 0.45-micron filter (Millipore) to remove eukaryotic- and bacterial cell-sized particles. The filtrate enriched in viral particles was treated with DNase and RNase to digest unprotected nucleic acid for 60 min. Remaining total nucleic acid was then isolated using QIAamp Viral RNA Mini Kit according to manufacturer's protocol. The libraries were then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the Illumina NovaSeq 6,000 with 250 bases paired ends with dual barcoding for each pool. For bioinformatics analysis, the generated 250 bp paired-end reads were debarcoded using vendor software from Illumina.