Liu Yusheng, Lu Falong, Wang Jiaqiang
College of Life Science, Northeast Forestry University, Harbin, China.
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2024;2723:215-232. doi: 10.1007/978-1-0716-3481-3_13.
Poly(A) tails are added to most eukaryotic mRNA and have essential regulatory functions. However, due to its homopolymeric nature, the sequence information in poly(A) tails is challenging to obtain in transcriptome measurement studies. In this chapter, we describe the detailed procedures of poly(A) inclusive full-length RNA isoform-sequencing (PAIso-seq), a method that can measure transcriptome-wide poly(A) tails from as low as nanogram level of total RNA based on the PacBio HiFi sequencing platform. The accurate length and base composition of poly(A) tails can be obtained along with the full-length cDNA.
多聚腺苷酸（Poly(A)）尾巴会添加到大多数真核生物mRNA上，并具有重要的调控功能。然而，由于其均聚物性质，在转录组测量研究中获取多聚腺苷酸尾巴中的序列信息具有挑战性。在本章中，我们描述了多聚腺苷酸包含的全长RNA异构体测序（PAIso-seq）的详细步骤，这是一种基于PacBio HiFi测序平台，能够从低至纳克级别的总RNA中测量全转录组范围多聚腺苷酸尾巴的方法。多聚腺苷酸尾巴的准确长度和碱基组成可与全长cDNA一同获得。
