Journal
GENOME BIOLOGY
Volume 15, Issue 11, Pages -Publisher
BIOMED CENTRAL LTD
DOI: 10.1186/s13059-014-0519-7
Keywords
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Funding
- National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services [HHSN272200900018C, HHSN272200900049C]
- National Institutes of Health, Office of Director, Innovator [DP2OD06514]
- NATIONAL HUMAN GENOME RESEARCH INSTITUTE [U01HG007480] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [U19AI115589, U19AI110818, R01AI104621] Funding Source: NIH RePORTER
- OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH [DP2OD006514] Funding Source: NIH RePORTER
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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.
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