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Data analysis pipeline for RNA-seq experiments: From differential expression to cryptic splicing

机译:RNA-seq实验的数据分析管道:从差异表达到隐秘剪接

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摘要

RNA sequencing (RNA-seq) is a high throughput technology that provides unique insights into the transcriptome. It has a wide variety of applications in quantifying genes/isoforms, detecting non-coding RNA, alternative splicing, and splice junctions. It is extremely important to comprehend the entire transcriptome for a thorough understanding of the cellular system. Several RNA-seq analysis pipelines are proposed to date. However, no single analysis pipeline can capture dynamics of the entire transcriptome. Here, we compile and present a robust and most commonly used analytical pipelines covering entire spectrum of transcriptome analysis, including quality checks, aligning reads, differential gene/transcript expression analysis, cryptic splicing events discovery, and visualization. Challenges, critical parameters, and possible downstream functional analysis pipelines associated with each step are highlighted and discussed. This provides a comprehensive understanding of state-of-the-art RNA-seq analysis pipeline and a greater understanding of the transcriptome.
机译:RNA测序(RNA-seq)是一项高通量技术,可提供对转录组的独特见解。它在定量基因/同工型,检测非编码RNA,选择性剪接和剪接处中具有广泛的应用。理解整个转录组对于彻底了解细胞系统非常重要。迄今为止,已提出了几种RNA-seq分析管道。但是,没有任何一个分析管道可以捕获整个转录组的动态。在这里,我们编译并展示了一个健壮且最常用的分析管道,涵盖了转录组分析的整个范围,包括质量检查,比对读取,差异基因/转录表达分析,隐秘剪接事件发现和可视化。与每个步骤相关的挑战,关键参数以及可能的下游功能分析流程都将得到重点介绍和讨论。这提供了对最新的RNA-seq分析流程的全面了解,并对转录组有了更深入的了解。

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