Enhanced Methods for Unbiased Deep Sequencing of Lassa and Ebola RNA Viruses from Clinical and Biological Samples.
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Date
2014
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Publisher
Genome Biology
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.