Genomics & Epigenomics Shared Resource, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC, USA.
Methods Mol Biol. 2024;2822:245-262. doi: 10.1007/978-1-0716-3918-4_17.
RNA sequencing (RNA-Seq) has emerged as a powerful and versatile tool for the comprehensive analysis of transcriptomes and has been widely used to investigate gene expression, copy number variation, alternative splicing, and novel transcript discovery. This chapter outlines the methodology for conducting short-read RNA-Seq, starting from RNA enrichment to library preparation and sequencing. Throughout the chapter, practical tips and best practices are provided to guide researchers in order to optimize each step of the RNA-Seq workflow. Multiple quality control steps throughout the workflow that are critical to obtain high-quality RNA-Seq data are also discussed.
RNA 测序 (RNA-Seq) 已经成为一种强大且多功能的转录组综合分析工具，广泛用于研究基因表达、拷贝数变异、可变剪接和新转录本的发现。本章概述了进行短读长 RNA-Seq 的方法学，从 RNA 富集到文库制备和测序。在本章中，提供了实用技巧和最佳实践，以指导研究人员优化 RNA-Seq 工作流程的每个步骤。还讨论了贯穿工作流程的多个关键质量控制步骤，以获得高质量的 RNA-Seq 数据。
