CattleFeces-Viromesh
Viral metagenome in feces from cattle in China
In 2015, 15 fecal samples were collected from cattle in Shanghai.The samples were resuspended in 10 volumes of phosphate-buffered saline (PBS) and vigorously vortexed for 5 min. Samples were pooled into a single pool. Five hundred microliters was collected after centrifugation (10 min, 15,000  g) and filtered through a 0.45-m filter (Millipore) to remove eukaryotic and bacterial cell-sized particles. The filtrate enriched in viral particles was treated with a mixture of DNases (Turbo DNase from Ambion, Baseline-ZERO from Epicentre, and benzonase from Novagen) and RNase (Fermentas) to digest unprotected nucleic acid at 37C for 90 min. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit (Qiagen) according to manufacturer's protocol. Viral cDNA synthesis was performed using SuperScript III reverse transcriptase (Invitrogen) according to the manufactures instructions. The second strand of cDNA synthesis was performed by incubation of reverse transcribed (RT) products with Klenow Fragment DNA polymerase (New England Biolabs). Library was 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.
CattleFeces-Viromesh
SUB4139002
CattleFeces-Viromesh