Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples
Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples
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DOI：
10.1186/s13059-014-0519-7
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发表时间：
2014-01-01
期刊：
影响因子：
12.3
通讯作者：
Sabeti, Pardis C.
中科院分区：
文献类型：
--
作者：
Matranga, Christian B.;Andersen, Kristian G.;Sabeti, Pardis C.
我们开发了一种强大的 RNA 测序方法，用于生成完整的从头组装，并在临床和生物样本中对拉沙病毒和埃博拉病毒基因组进行宿主内变异调用。我们的方法使用基于 RNase H 的靶向消化来去除污染的聚 (rA) 载体和核糖体 RNA。这一去除步骤提高了无偏总 RNA 测序文库中的数据质量和信息读数的数量。我们还开发了混合选择方案以进一步丰富测序文库的病毒内容。这些方案能够对拉沙病毒和埃博拉病毒进行快速深度测序，并广泛适用于其他病毒基因组学研究。
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.