Explore in-depth RNA sequencing results
The Iso-Seq method enables in-depth analysis of cDNA sequencing across the entire transcriptome or targeted genes. The resulting full-length mRNA transcript sequences can be easily obtained through a simple analysis workflow. Iso-Seq analysis capabilities also include the discovery of novel transcripts and genes, the identification of fusion genes, and the annotation of isoforms and alternative splicing events.
SMRT Analysis algorithms for RNA sequencing analysis
The Iso-Seq method
Using the long reads generated by Single Molecule, Real-Time (SMRT) Sequencing, the Iso-Seq method provides reads that span entire transcript isoforms, from the 5′ end to the 3′ polyA-tail. Generation of accurate, full-length transcript sequences greatly simplifies analysis by eliminating the need for transcript reconstruction to infer isoforms using error-prone assembly of short RNA-seq reads.
SMRT Analysis applications for RNA sequencing analysis
This application enables functional characterization of transcripts and splice variants for data generated with SMRT Sequencing technology.
Iso-Seq analysis runs either in de novo or reference-based mode.
It includes three main steps:
- Classify: Extract reads of insert from PacBio Systems (or SMRT Cell) runs; remove cDNA primers and poly-As; then classify the reads of insert into chimeric or non-chimeric, and full-length or non-full-length reads.
- Cluster: Predict de novo consensus isoforms of transcripts from the classified reads using the Iteratively Clustering and Error Correction (ICE) algorithm.
- Map: Using GMAP, align the classified reads and predicted consensus isoforms to the user-specific reference sequence.
Contact us to learn more about how this innovative software can support your research efforts.