Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples

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Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples

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Title: Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples
Author: Matranga, Christian B; Andersen, Kristian G; Winnicki, Sarah; Busby, Michele; Gladden, Adrianne D; Tewhey, Ryan; Stremlau, Matthew; Berlin, Aaron; Gire, Stephen K; England, Eleina; Moses, Lina M; Mikkelsen, Tarjei S; Odia, Ikponmwonsa; Ehiane, Philomena E; Folarin, Onikepe; Goba, Augustine; Kahn, S Humarr; Grant, Donald S; Honko, Anna; Hensley, Lisa; Happi, Christian; Garry, Robert F; Malboeuf, Christine M; Birren, Bruce W; Gnirke, Andreas; Levin, Joshua Z; Sabeti, Pardis C

Note: Order does not necessarily reflect citation order of authors.

Citation: Matranga, C. B., K. G. Andersen, S. Winnicki, M. Busby, A. D. Gladden, R. Tewhey, M. Stremlau, et al. 2014. “Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples.” Genome Biology 15 (11): 519. doi:10.1186/s13059-014-0519-7. http://dx.doi.org/10.1186/s13059-014-0519-7.
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Abstract: We have developed a robust RNA sequencing method for generating complete de novo assemblies with intra-host variant calls of Lassa and Ebola virus genomes in clinical and biological samples. Our method uses targeted RNase H-based digestion to remove contaminating poly(rA) carrier and ribosomal RNA. This depletion step improves both the quality of data and quantity of informative reads in unbiased total RNA sequencing libraries. We have also developed a hybrid-selection protocol to further enrich the viral content of sequencing libraries. These protocols have enabled rapid deep sequencing of both Lassa and Ebola virus and are broadly applicable to other viral genomics studies. Electronic supplementary material The online version of this article (doi:10.1186/s13059-014-0519-7) contains supplementary material, which is available to authorized users.
Published Version: doi:10.1186/s13059-014-0519-7
Other Sources: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4262991/pdf/
Terms of Use: This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA
Citable link to this page: http://nrs.harvard.edu/urn-3:HUL.InstRepos:13581031
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