June 1, 2021

A complete solution for full-length transcript sequencing using the PacBio Sequel II System

Author(s): Underwood, Jason and Galvin, Brendan and Hon, Ting and Tseng, Elizabeth

Long read mRNA sequencing methods such as PacBio’s Iso-Seq method offers high-throughput transcriptome profiling in prokaryotic and eukaryotic cells. By avoiding the transcript assembly problem and instead sequencing full-length cDNA, Iso-Seq has emerged as the most reliable technology for annotating isoforms and, in turn, improving proteome predictions in a wide variety of organisms. Improvements in library preparation, sequencing throughput, and bioinformatics has enabled the Iso-Seq method to be complete solution for transcript characterization. The Iso-Seq Express kit is a one-day library prep requiring 60-300 ng of total RNA. The PacBio Sequel II system produces 4-5 million full-length reads, sufficient to profile a whole human transcriptome. Finally, the SQANTI2 software is a powerful tool for categorizing the complex isoforms against reference annotations, while also incorporating orthogonal information such as CAGE peak data, public RNA-seq junction data, and ORF predictions.

Organization: PacBio
Year: 2020

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