International Institute of Molecular and Cell Biology, Warsaw, Poland.
Institute of Biochemistry and Biophysics, Warsaw, Poland.
Methods Mol Biol. 2024;2723:193-214. doi: 10.1007/978-1-0716-3481-3_12.
There are multiple methods for studying deadenylation, either in vitro or in vivo, which allow for observation of mRNA abundance or poly(A) tail dynamics. However, direct RNA sequencing using the Oxford Nanopore Technologies (ONT) platform makes it possible to conduct transcriptome-wide analyses at the single-molecule level without the PCR bias introduced by other methods. In this chapter, we provide a protocol to measure both RNA levels and poly(A)-tail lengths in the yeast Saccharomyces cerevisiae using ONT.
有多种方法可用于研究脱腺苷酸化，无论是在体外还是体内，这些方法都可以观察到 mRNA 丰度或 poly(A)尾动力学。然而，使用 Oxford Nanopore Technologies（ONT）平台进行直接 RNA 测序使得在无需其他方法引入的 PCR 偏倚的情况下，在单分子水平上进行全转录组分析成为可能。在本章中，我们提供了一种使用 ONT 测量酵母酿酒酵母中 RNA 水平和 poly(A)尾长度的方案。
