Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
Division of Nutritional Sciences, Cornell University, Ithaca, NY, USA.
Methods Mol Biol. 2021;2252:221-237. doi: 10.1007/978-1-0716-1150-0_10.
Ribosome profiling is a powerful technique that enables researchers to monitor translational events across the transcriptome. It provides a snapshot of ribosome positions and density across the transcriptome at a sub-codon resolution. Here we describe the whole procedure of profiling ribosome footprints in mammalian cells. Two methods for Ribo-seq library construction are introduced, and their advantages and disadvantages are compared. There is a room for further improvement of Ribo-seq in terms of the amount of starting material, the duration of library construction, and the resolution of sequencing results.
核糖体图谱分析是一种强大的技术，使研究人员能够监测转录组中的翻译事件。它提供了在亚密码子分辨率下跨转录组的核糖体位置和密度的快照。在这里，我们描述了在哺乳动物细胞中进行核糖体足迹图谱分析的整个过程。介绍了两种 Ribo-seq 文库构建方法，并比较了它们的优缺点。在起始材料的数量、文库构建的持续时间和测序结果的分辨率方面，Ribo-seq 还有进一步改进的空间。
