Author: Piewbang, Chutchai; Rungsipipat, Anudep; Poovorawan, Yong; Techangamsuwan, Somporn
Title: Cross-sectional investigation and risk factor analysis of community-acquired and hospital-associated canine viral infectious respiratory disease complex Document date: 2019_11_14
ID: r5oefcrj_13
Snippet: Viral nucleic acids were extracted from the NS or OS samples (above) using a Viral Nucleic Acid Extraction Kit II (GeneAid, Xizhi, Taiwan) according to the manufacturer's protocol, and then quantified and qualified using Nanodrop® Lite (Thermo Fisher Scientific Inc., Massachusetts, U.S.A.) at an absorbance of 260 and 280 nm. The extracted nucleic acid was divided into two aliquots, one for two-step RT-PCR for RNA viruses (CIV, CPIV, CDV, and CRC.....
Document: Viral nucleic acids were extracted from the NS or OS samples (above) using a Viral Nucleic Acid Extraction Kit II (GeneAid, Xizhi, Taiwan) according to the manufacturer's protocol, and then quantified and qualified using Nanodrop® Lite (Thermo Fisher Scientific Inc., Massachusetts, U.S.A.) at an absorbance of 260 and 280 nm. The extracted nucleic acid was divided into two aliquots, one for two-step RT-PCR for RNA viruses (CIV, CPIV, CDV, and CRCoV) and the other for direct PCR for DNA viruses (CAdV-2 and CaHV-1). For the first stage RT-PCR assay, 100 ng of total RNA was used as the template to generate complementary DNA (cDNA) using the Omniscript® Reverse Transcription Kit (Qiagen GmbH, Hilden, Germany). The cDNA and DNA were kept at -20 C until used for subsequent PCR amplifications.
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