Blood virome of cattle from forest region revealed diverse small circular ssDNA viral genomes
During 2015, 80 bovine blood samples were collected from a forested region in Northeast China. These samples were collected as part of a survey for arthropod-borne viral infection in cattle. Library preparation and computational analysis were performed as previously described. Briefly, 80 blood samples with anticoagulants were pooled into five pools which included 16 blood samples each. Supernatants were then collected after centrifugation (10 min, 13,000 ¡Á g). Plasma pools were filtered through a 0.45-mm filter (Millipore) to remove eukaryotic- and bacterial cell-sized particles, and 200 uL of each pool was then subjected to a mixture of nuclease enzymes to reduce the concentration of free (non-viral encapsidated) nucleic acids. Remaining total nucleic acid was then isolated using QiaAmp Mini Viral RNA kit (Qiagen) according to manufacturer's protocol. Viral cDNA synthesis was performed using SuperScript III reverse transcriptase _(Invitrogen) according to the manufacture¡¯s instructions. The second strand of cDNA synthesis _was performed by incubation of reverse transcribed (RT) products with Klenow Fragment _DNA polymerase (New England Biolabs). Five libraries were then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the MiSeqIllumina platform with 250 bases paired ends with dual barcoding for each pool. Bioinformatics analysis was performed according to a previous study.
SUB3100280
Cattleblood-Virome