PAIso-seq

PAIso-seq quantifies transcriptome-wide poly(A) tail length and composition from full-length cDNA using PacBio HiFi sequencing to investigate post-transcriptional regulation of mRNA.

Key Features:

• High Sensitivity: Detects poly(A) tails from subnanogram total RNA inputs, enabling analysis of single mammalian oocytes (~0.5 ng total RNA).
• Comprehensive Analysis: Provides transcriptome-wide measurement of poly(A) tail length and identification of non-A residues while capturing full-length cDNA for isoform-level information.
• PacBio Sequencing Integration: Leverages PacBio sequencing technology in HiFi mode to produce high-fidelity long reads for precise poly(A) tail characterization.
• Detailed Protocol and Bioinformatics Pipeline: Includes a library preparation protocol for single or bulk oocyte samples and a bioinformatic pipeline for processing raw sequencing data through initial analysis.

Scientific Applications:

• Low-input sample analysis: Enables poly(A) tail and transcriptome profiling from precious in vivo samples with limited RNA, including single oocytes.
• Post-transcriptional regulation studies: Supports investigation of mRNA export, stability, and translation by providing tail length and composition data.
• Isoform-resolved transcriptomics: Captures full-length cDNA to facilitate isoform-level analyses and transcriptome-wide studies of poly(A) tail dynamics.

Methodology:

Initial data analysis uses the provided bioinformatics pipeline to process raw PacBio HiFi data and can be completed within eight hours.