title: Identification and characterization of a novel recombinant porcine astrovirus from pigs in Anhui, China:lib_Anhuipig
design: In 2018, 215 pig feces samples were collected from a pig farm in Anhui Province, China. All samples were collected using disposable 1.5ml eppendorf (EP) tubes and shipped on dry ice. Fecal samples were resuspended individually in 1ml of phosphate-buffered saline (PBS) and vigorously vortexed for 5 min. After centrifugation at 12,000 rpm for 10 min, the 215 feces sample supernatants were then collected by pipette and were stored at -80 before nucleic acid extraction.20 feces sample supernatants were randomly choose to generate one pool, the total 500l mixed supernatant was firstly filtered through a 0.45-m filter (Millipore) to remove eukaryotic and bacterial cell-sized particles, then the viral particle enrichment filtrate were treated with DNase and RNase to digest unprotected non-viral nucleic acid at 37 C for 60 min. The total nucleic acid was then extracted using QiaAmp Mini Viral RNA kit (Qiagen) according to manufacturers protocol. Libraries were constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the MiSeq Illumina platform with 250 bases paired ends with dual barcoding for each pool. Paired-end reads of 250 bp generated by MiSeq were debarcoded using vendor software from Illumina. An in-house analysis pipeline running on a 32-node Linux cluster was used to process the data.