Identification and full-genome characterization of novel circovirus in giant pandas (Ailuropoda melanoleuca)

In 2019, to investigate the virome of giant pandas, 12 blood samples were collected in China. All samples were collected by disposable materials. Each sample was resuspended in ten volumes of phosphate buffered saline and vigorously vortexed for 5 min. The supernatants were then collected after centrifugation. 500 ul of each supernatant was filtered through a 0.45 um filter to remove eukaryotic and bacterial cell sized particles. The filtrates enriched in viral particles were treated with DNase and RNase to digest unprotected nucleic acid for 60 min. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit according to manufacturer protocol. One library was then constructed using Nextera XT DNA Sample Preparation Kit and sequenced using the Illumina NovaSeq 6000 platform with 250 bases paired ends with dual barcoding for each library. For bioinformatics analysis, paired end reads of 250 bp generated by Illumina MiSeq or NovaSeq platform were debarcoded using vendor software from Illumina.