Rna-seq of  Rattus norvegicus :  viral rna pool of throat swab and anal swab samples collected from 15-20 rodents in Tunchang, Nandian ,China
Swab sample were passed through 0.45m filters (Millipore, Darmstadt, Germany) to remove eukaryotic and bacterium-sized particles. The filtrate was ultracentrifuged at 30,000g, 4 for 3h. The precipitate collected from 28 pool samples were re-suspended with Hanks balanced salt solution and digested with DNase (Applied Biosystems/USA) and RNase One (Promega/USA) to remove unprotected nucleic acids. The viral DNA and RNA was extracted using a QIAamp Viral RNA Mini Kit (QIAGEN/USA) according to the manufacturers instructions. The viral first-strand cDNA was generated using Superscript III Reverse Transcriptase (Invitrogen/USA) and primer K-8N, as described previously(22). Convert cDNA to dsDNA by incubating cDNA with Klenow fragment (NEB, Ipswich, MA, USA) at 37C for 1 h. Sequence-independent PCR amplification products were purified with a MinElute PCR Purification Kit (Qiagen) and subjected to magnetic bead sorting. Viral nucleic acid libraries constructed by preprocessing such as sonication were analyzed using an Illumina HiSeq2500 sequencer (Illumina, USA).