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Use Of S1 Nuclease In Deep Sequencing For Detection Of Double-Stranded Rna Viruses

JOURNAL OF VETERINARY MEDICAL SCIENCE(2015)

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摘要
Metagenomic approach using next-generation DNA sequencing has facilitated the detection of many pathogenic viruses from fecal samples. However, in many cases, majority of the detected sequences originate from the host genome and bacterial flora in the gut. Here, to improve efficiency of the detection of double-stranded (ds) RNA viruses from samples, we evaluated the applicability of Si nuclease on deep sequencing. Treating total RNA with Si nuclease resulted in 1.5-28.4- and 10.1-208.9-fold increases in sequence reads of group A rotavirus in fecal and viral culture samples, respectively. Moreover, increasing coverage of mapping to reference sequences allowed for sufficient genotyping using analytical software. These results suggest that library construction using Si nuclease is useful for deep sequencing in the detection of dsRNA viruses.
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关键词
deep sequencing, double-stranded RNA viruses, group a rotavirus, next-generation sequencer, S1 nuclease
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