Translational Oncology Group, Galicia Sur Health Research Institute (IIS Galicia Sur), SERGAS-UVIGO, Vigo, Spain.
Methods Mol Biol. 2025;2876:151-162. doi: 10.1007/978-1-0716-4252-8_10.
RNA-sequencing (RNA-seq) has become the method of choice for whole transcriptome analysis, as it enables profiling of a wide range of RNA molecules and the always evolving bioinformatic pipelines allow the extraction of diverse biological information from a single sample. However, there is not a gold-standard RNA-seq protocol nor an optimal bioinformatic pipeline defined, which can challenge new users. The aim of this chapter is to describe the basic RNA-sequencing pipeline, from RNA extraction and library preparation to the bioinformatic preprocessing and downstream analyses. The steps are oriented to the transcriptome analysis of the skin of TR-deficient mice, but the protocol can be easily translated to other tissue types or organisms.
RNA 测序（RNA-seq）已成为全转录组分析的首选方法，因为它能够对广泛的 RNA 分子进行分析，并且不断发展的生物信息学管道允许从单个样本中提取多种生物学信息。然而，目前还没有一个金标准的 RNA-seq 方案或定义明确的最佳生物信息学管道，这可能会给新用户带来挑战。本章的目的是描述基本的 RNA-seq 管道，从 RNA 提取和文库制备到生物信息学预处理和下游分析。这些步骤面向 TR 缺陷型小鼠皮肤的转录组分析，但该方案可以很容易地转化为其他组织类型或生物体。
