Full-Genome Deep Sequencing and Phylogenetic Analysis of Novel Human Betacoronavirus
Matthew Cotten, Tommy T. Lam, Simon J. Watson, Anne L. Palser, Velislava Petrova, Paul Grant, Oliver G. Pybus, Andrew Rambaut, Yi Guan, Deenan Pillay, Paul Kellam , and Eleni Nastouli
Author affiliations: Wellcome Trust Sanger Institute, Hinxton, UK (M. Cotten, S.J. Watson, A.L. Palser, V. Petrova, P. Kellam); University of Oxford, Oxford, UK (T.T. Lam, O.G. Pybus); University College London, London, UK (D. Pillay, P. Kellam); University College London Hospitals,; London (P.Grant, E. Nastouli); University of Edinburgh, Edinburgh, Scotland, UK (A. Rambaut); Fogarty International Center–National Institutes for Health, Bethesda, Maryland, USA (A. Rambaut); The University of Hong Kong, Hong Kong (Y. Guan)
Figure 1. . . . A) Primers designed for reverse transcription and overlapping PCR amplification of the novel coronavirus (CoV). Dots indicate the predicted binding site of each primer along the EMC/2012 genome (x-axis). Gray bars indicate predicted amplicon lengths. Amplicon numbers are indicated beside each set of products. B) PCR products (3 µL of a 25-µL reaction) were resolved by electrophoresis on a 0.6% agarose gel and visualized by ethidium bromide staining. Lane M is the molecular weight marker (sizes indicated at left), Lanes 1–15 show the products of the amplicons depicted in Panel A. Lane C is the reagent PCR control.
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