核糖核酸
RNA序列
计算生物学
转录组
聚腺苷酸
生物
非编码RNA
小RNA
转录后修饰
RNA编辑
基因
基因表达
遗传学
作者
Kimberly R. Kukurba,Stephen B. Montgomery
出处
期刊:CSH Protocols
[Cold Spring Harbor Laboratory]
日期:2015-04-13
卷期号:2015 (11): pdb.top084970-pdb.top084970
被引量:711
标识
DOI:10.1101/pdb.top084970
摘要
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.
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