Iso-Seq

Isoform sequencing (Iso-Seq) is a type of RNA sequencing (RNA-Seq) method that uses Single Molecular, Real-Time (SMRT^®) long-read technology to sequence contiguous, full-length transcripts. This method addresses the errors that can occur using short-read approaches and captures the transcript start, polyadenylation, and splice sites from a single read. Applications include better-annotated genomes, detection of gene fusion, and discovery of novel isoforms.

Instead of the RSII, our Iso-Seq service leverages the PacBio Sequel platforms to enhance Iso-Seq capabilities. Additionally, we have optimized the process to exceed output and read-length benchmarks set by PacBio to ensure you receive high-quality results.

Contact our team to discuss how Iso-Seq on the new PacBio system can improve your NGS projects.

What is Iso-Seq?

The Iso-Seq method generates full-length cDNA sequences and analyzes full-length transcript isoforms, which can be used in RNA sequencing (RNA-Seq) to:

See alternative start and end sites
Characterize splicing events
Find fusion transcripts
Identify allele-specific isoforms
Discover non-coding RNAs
Profile expression at isoform resolution
Predict full-length open read frames and more

What is the output of Iso-Seq?

The Iso-Seq method and bioinformatics workflow outputs high-quality, full-length transcript sequences of 10kb or longer. The high accuracy of high-fidelity (HiFi) reads ensure that SNPs can be identified, as well as UMIs and barcodes used for single-cell studies.

Iso-Seq delivers full-length transcript isoforms with no assembly required. Unlike short-read RNA-Seq methods, Iso-Seq method reads span the whole transcript, capturing even untranslated regions.

What is Iso-Seq analysis using PacBio technology?

Iso-Seq analysis empowers you to discover novel transcripts and genes, identify fusion genes, and annotate isoforms and alternative splicing events. Iso-Seq analysis uses highly accurate long reads (HiFi reads) generated with the CCS algorithm to cluster and generate full-length, high-quality transcript isoforms.

What is the depth of Iso-Seq?

The read depth of Iso-Seq is 1.6M full length transcripts.

TECHNICAL DETAILS

Full-Length RNA-Seq: A Novel Method to Assess Sequence Integrity for RNA Therapeutics

Strict quality control is required to maintain the integrity of manufactured products for RNA therapies, but current assays often present limitations. Learn how the novel full-length RNA-Seq approach from Azenta allows you to preserve the entire length of and effectively sequence the poly(A) tails of your mRNA products.

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Isoform Sequencing on PacBio Sequel – Maximizing Output and Accuracy

Learn how Azenta has further optimized PacBio Sequel capabilities to exceed PacBio’s benchmarks in output and read length.

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Article: Which RNA-Seq Technique Should I Use?

With so many RNA sequencing assay types to choose from, how do you know which is best suited for your NGS project? In this article, we discuss the most common RNA-Seq approaches and what factors to consider when selecting the right one.

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Blog: PacBio HiFi Sequencing for Understanding Human Health and Disease

At the Azenta Life Sciences and PacBio virtual symposium titled, Decoding the Complexity of Human Health – A HiFi Vision, speakers from leading research institutions and biotech discussed the role of high-fidelity, long-read sequencing in understanding human health and diseases. In this blog post, we recap the presentations and highlight the themes that emerged from the virtual event.

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Features & Benefits

Optimized workflows deliver best output and accuracy when compared to PacBio benchmarks

Dedicated support from experienced Azenta scientists

Lower input material requirements when compared to older technologies

Cost-effective with fast turnaround times

NGS Platforms

Azenta is a qualified service provider for Illumina and PacBio NGS platforms. For information on our NGS platforms as well as recommended configurations for your projects, please visit the NGS Platforms page.

Technical Resources

Deliverables

All customers receive their raw data as BAM files. Additional data analysis including: annotation and full-length non-chimeric reads are available. Other custom reports are also available on request.