RNA Sequencing and Analysis.
RNA Sequencing and Analysis.
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DOI：
10.1101/pdb.top084970
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发表时间：
2015-04-13
影响因子：
--
通讯作者：
Montgomery SB
中科院分区：
文献类型：
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作者：
Kukurba KR;Montgomery SB
RNA测序（RNA-Seq）使用高通量测序方法的能力来提供对细胞转录组的深入了解。与以前的基于桑格测序和微阵列的方法相比，RNA-Seq提供了更高的覆盖率和转录组动态性质的更高分辨率。除了定量基因表达外，RNA-Seq生成的数据还有助于发现新的转录本、鉴定可变剪接基因和检测等位基因特异性表达。RNA-Seq工作流程的最新进展，从样品制备到文库构建再到数据分析，使研究人员能够进一步阐明转录的功能复杂性。除了聚腺苷酸化信使RNA（mRNA）转录物外，RNA-Seq还可用于研究不同的RNA群体，包括总RNA、前mRNA和非编码RNA，如microRNA和长ncRNA。本文介绍了RNA-Seq方法，包括应用，实验设计和技术挑战。
RNA sequencing (RNA-Seq) uses the capabilities of high-throughput sequencing methods to provide insight into the transcriptome of a cell. Compared to previous Sanger sequencing- and microarray-based methods, RNA-Seq provides far higher coverage and greater resolution of the dynamic nature of the transcriptome. Beyond quantifying gene expression, the data generated by RNA-Seq facilitate the discovery of novel transcripts, identification of alternatively spliced genes, and detection of allele-specific expression. Recent advances in the RNA-Seq workflow, from sample preparation to library construction to data analysis, have enabled researchers to further elucidate the functional complexity of the transcription. In addition to polyadenylated messenger RNA (mRNA) transcripts, RNA-Seq can be applied to investigate different populations of RNA, including total RNA, pre-mRNA, and noncoding RNA, such as microRNA and long ncRNA. This article provides an introduction to RNA-Seq methods, including applications, experimental design, and technical challenges.