Sequencing of Transcriptome-Wide Poly(A) Tails by PAIso-seq

Yusheng Liu; Falong Lu; Jiaqiang Wang

doi:10.1007/978-1-0716-3481-3_13

Sequencing of Transcriptome-Wide Poly(A) Tails by PAIso-seq

Methods Mol Biol. 2024:2723:215-232. doi: 10.1007/978-1-0716-3481-3_13.

Authors

Yusheng Liu¹, Falong Lu^{2

3}, Jiaqiang Wang⁴

Affiliations

¹ College of Life Science, Northeast Forestry University, Harbin, China.
² State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China. [email protected].
³ University of Chinese Academy of Sciences, Beijing, China. [email protected].
⁴ College of Life Science, Northeast Agricultural University, Harbin, China. [email protected].

PMID: 37824073
DOI: 10.1007/978-1-0716-3481-3_13

Abstract

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.

Keywords: Full-length RNA isoform; Mixed tailing; PAIso-seq; Poly(A) tail; Transcriptome-wide.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA, Complementary
High-Throughput Nucleotide Sequencing
Poly A / genetics
Poly A / metabolism
Polyadenylation*
RNA, Messenger / genetics
RNA, Messenger / metabolism
Sequence Analysis, RNA / methods
Transcriptome*

Substances

DNA, Complementary
RNA, Messenger
Poly A