Transcriptomics is a crucial field in biological research, dedicated to exploring gene expression and its regulatory mechanisms. At its core lies the comprehensive analysis of gene expression patterns across diverse tissues, cells, or experimental conditions. Traditional short-read and long-RNA sequencing technologies, such as those offered by Illumina, face limitations due to fragmentation. This fragmentation prevents the complete coverage of full-length transcripts, leading to incomplete sequence splicing and inaccurate gene annotation. In contrast, full-length transcript sequencing provides comprehensive information about transcripts, including the 5′ and 3′ exons, introns, and poly-A tails. This detailed information enables a more in-depth understanding of gene expression and regulatory mechanisms.

Iso-Seq technology is a full-length transcriptome sequencing method based on single molecule real-time (SMRT) sequencing, which was developed by PacBio company. By synthesizing the full-length cDNA library, this technology can directly capture the complete transcript sequence by using its super-long reading ability (the average length can reach 10-15 kb) without relying on the reference genome. Iso-Seq technology has significant advantages in sequencing full-length transcripts, such as:

High Precision in detection: Iso-Seq showcases remarkable accuracy in identifying splicing events, transcript isoforms, fusion genes, and non – coding RNAs. This precision provides researchers with a more nuanced and accurate view of the transcriptome, enabling a deeper understanding of gene function and regulation.

Comprehensive Transcript Coverage: Covering the complete transcript from 5′ end to 3′ end, including intron and poly-A tail, which is helpful to analyze the complex transcriptome structure.

Versatility in application: It is suitable for full-length transcriptome sequencing of species without reference genome, and supports parallel analysis of multi-tissue samples.

Enhanced effects through interfration: Combining with other sequencing technologies (such as RNA-seq) can further improve the parsing ability of transcriptome data.

PacBio is the main promoter and leader of Iso-Seq technology. Sequel II series, its third-generation sequencing platform, has significantly improved the accuracy and consistency of full-length transcripts through optimized HiFi sequencing technology. PacBio’s Iso-Seq technology is not only widely used in academic research, but also used for genome annotation and transcriptome analysis of plants, animals and humans.

In addition, PacBio has developed a variety of supporting tools and software (such as SMRT Link, IsoCon, TAMA, etc.) to provide us with a complete solution from sample preparation to data analysis.
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Full-Length Transcripts Sequencing (Iso-Seq)
PacBio SMRT Sequencing