viral metagenome Metagenome

From April to May in spring 2018 for viral metagenomic analysis 123 fecal samples of diarrhea piglets were collected from farms in Shanghai City China All piglets were under one month of age All samples were collected by disposable materials Fecal samples were resuspended with 10 volumes of phosphate buffered saline PBS and vigorously rotated for 5 minutes After centrifugation 10min 15000 g the supernatant was placed in a centrifuge tube 500 of the mixed supernatant was filtered through a microwell filter in order to remove particles such as bacteria and eukaryotes Fecal samples were treated at 37 for 90 minutes to digest free non-viral nucleic acids and the filtrate that had been enriched in virus particles was treated with a mixture of DNase and RNase A QIAamp Viral RNA Mini Kit QIAGEN was used to extract nucleic acid according to the manufacturer's instruction 89 libraries were then constructed using Nextera XT DNA Sample Preparation Kit Illumina and sequenced using the Illumina Miseq platform with 250 bases paired ends with double barcodes for each library For bioinformatics analysis paired-end reads of 250 bp generated by MiSeq were debarcoded using vendor software from Illumina