Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing

RNA viruses are known for a high mutation rate and rapid genomic evolution. As such an RNA virus population does not consist of a single genotype but is rather a collection of individual viruses with closely related genotypes—a quasispecies, which can be analyzed by next-generation sequencing (NGS)....

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Autores principales: Keep, Sarah M., Bickerton, Erica, Britton, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2014
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7122900/
https://www.ncbi.nlm.nih.gov/pubmed/25720476
http://dx.doi.org/10.1007/978-1-4939-2438-7_11
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author Keep, Sarah M.
Bickerton, Erica
Britton, Paul
author_facet Keep, Sarah M.
Bickerton, Erica
Britton, Paul
author_sort Keep, Sarah M.
collection PubMed
description RNA viruses are known for a high mutation rate and rapid genomic evolution. As such an RNA virus population does not consist of a single genotype but is rather a collection of individual viruses with closely related genotypes—a quasispecies, which can be analyzed by next-generation sequencing (NGS). This diversity of genotypes provides a mechanism in which a virus population can evolve and adapt to a changing environment. Sample preparation is vital for successful sequencing. The following protocol describes the process of generating a high-quality RNA preparation from IBV grown in embryonated eggs and then partially purified and concentrated through a 30 % sucrose cushion for NGS.
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spelling pubmed-71229002020-04-06 Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing Keep, Sarah M. Bickerton, Erica Britton, Paul Coronaviruses Article RNA viruses are known for a high mutation rate and rapid genomic evolution. As such an RNA virus population does not consist of a single genotype but is rather a collection of individual viruses with closely related genotypes—a quasispecies, which can be analyzed by next-generation sequencing (NGS). This diversity of genotypes provides a mechanism in which a virus population can evolve and adapt to a changing environment. Sample preparation is vital for successful sequencing. The following protocol describes the process of generating a high-quality RNA preparation from IBV grown in embryonated eggs and then partially purified and concentrated through a 30 % sucrose cushion for NGS. 2014-12-18 /pmc/articles/PMC7122900/ /pubmed/25720476 http://dx.doi.org/10.1007/978-1-4939-2438-7_11 Text en © Springer Science+Business Media New York 2015 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Article
Keep, Sarah M.
Bickerton, Erica
Britton, Paul
Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing
title Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing
title_full Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing
title_fullStr Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing
title_full_unstemmed Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing
title_short Partial Purification of IBV and Subsequent Isolation of Viral RNA for Next-Generation Sequencing
title_sort partial purification of ibv and subsequent isolation of viral rna for next-generation sequencing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7122900/
https://www.ncbi.nlm.nih.gov/pubmed/25720476
http://dx.doi.org/10.1007/978-1-4939-2438-7_11
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