使用RNA测序进行RNA定量分析

Quantitative RNA Analysis Using RNA-Seq.

作者信息

Myler Peter J, McDonald Jacqueline A, Alcolea Pedro J, Sur Aakash

机构信息

Center for Global Infectious Disease Research, Seattle Childrens Research Institute, 307 Westlake Ave N, Suite 500, Seattle, 98109-5219, WA, USA.

Department of Global Health, University of Washington, Seattle, 98195, WA, USA.

出版信息

Methods Mol Biol. 2019;1971:95-108. doi: 10.1007/978-1-4939-9210-2_4.

DOI:10.1007/978-1-4939-9210-2_4

PMID:30980299

Abstract

High-throughput sequencing of cDNA copies of mRNA (RNA-seq) provides a digital readout of mRNA levels over several orders of magnitude, as well as mapping the transcripts to the nucleotide level. Here we describe two different RNA-seq approaches, including one that exploits the 39-nucleotide mini-exon or spliced leader (SL) sequence found at the 5' end of all Leishmania (and other trypanosomatid) mRNAs.

摘要

信使核糖核酸（mRNA）的互补脱氧核糖核酸（cDNA）拷贝的高通量测序（RNA测序）可提供跨越多个数量级的mRNA水平数字读数，还能在核苷酸水平上对转录本进行定位。在此，我们描述两种不同的RNA测序方法，其中一种利用了在所有利什曼原虫（以及其他锥虫）mRNA 5'端发现的39个核苷酸的小外显子或剪接前导序列（SL）。