Lee-Liu Dasfne, Almonacid Leonardo I, Faunes Fernando, Melo Francisco, Larrain Juan
Center for Aging and Regeneration and Millennium Nucleus in Regenerative Biology, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Methods Mol Biol. 2012;917:293-317. doi: 10.1007/978-1-61779-992-1_18.
Next generation sequencing technologies may now be applied to the study of transcriptomics. RNA-Seq or RNA sequencing employs high-throughput sequencing of complementary DNA fragments delivering a transcriptional profile. In this chapter, we aim to provide a starting point for Xenopus researchers planning on starting an RNA-Seq transcriptomics study. We begin by providing a section on template isolation and library preparation. The next section comprises the main bioinformatics procedures that need to be performed for raw data processing, normalization, and differential gene expression. Finally, we have included a section on studying deep sequencing results in Xenopus, which offers general guidance as to what can be done in this model.
新一代测序技术现在可应用于转录组学研究。RNA测序（RNA-Seq）采用对互补DNA片段进行高通量测序，从而提供转录图谱。在本章中，我们旨在为计划开展RNA-Seq转录组学研究的非洲爪蟾研究人员提供一个起点。我们首先提供关于模板分离和文库制备的部分内容。下一部分包括对原始数据处理、标准化和差异基因表达进行分析所需的主要生物信息学程序。最后，我们纳入了一部分关于研究非洲爪蟾深度测序结果的内容，这部分针对在该模型中可以开展的研究提供了一般性指导。
