Next Generation Sequencing of Lyssaviruses

Denise A. Marston*, Lorraine M. McElhinney, Emma Wise, Richard J. Ellis, Conrad M. Freuling, Thomas F. Müller, Anthony R. Fooks

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

With the advent of Next Generation Sequencing (NGS) technologies, the ability to quickly generate large amounts of sequence data has revolutionized the genomics field. Most RNA viruses have relatively small genomes in comparison to other organisms and as such, would be expected to easily generate genome sequence data via NGS technologies. However, due to the relatively low abundance of viral RNA in relation to host RNA, RNA viruses have proved relatively difficult to sequence using NGS technologies. Here, we detail a simple, robust methodology, without the use of ultra-centrifugation, filtration or viral enrichment protocols, to prepare RNA from diagnostic clinical tissue samples, cell monolayers and tissue culture supernatant, for subsequent sequencing on the Roche 454 platform.

Original languageEnglish
Title of host publicationCurrent Laboratory Techniques in Rabies Diagnosis, Research and Prevention
PublisherElsevier Inc.
Pages171-183
Number of pages13
Volume1
ISBN (Electronic)9780128004654
ISBN (Print)9780128000144
DOIs
Publication statusPublished - 4 Aug 2014
Externally publishedYes

Bibliographical note

Funding Information:
This work was funded by the Department for Environment, Food and Rural Affairs (Defra), UK (grant ROAME SE0427), and by the EU FP7-funded Research Infrastructure Grant European Virus Archive (No. 19 228292).

Publisher Copyright:
© 2014 Crown Copyright and Elsevier Inc. All rights reserved.

Keywords

  • Genome Lyssavirus
  • Next Generation Sequencing (NGS)
  • Polymerase Chain Reaction (PCR)
  • RNA
  • Roche 454 Platform
  • TRIzol<sup>®</sup>

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