RNA sequencing with the Iso-Seq method
Reveal novel biology that other sequencing methods miss with full-length transcripts using the Iso-Seq™ method
Human transcriptome studies
Explore how changes in isoform usages contribute to the phenotypic differences between health and disease.
Plant + animal transcriptome studies
Advance breeding programs and basic research with full-length cDNA sequencing that does not require a reference genome.
Single-cell transcriptome studies
Visualize biology at an even higher resolution by capturing a comprehensive and complete transcript sequence from individual cells.
Introduction to RNA sequencing using the Iso-Seq method
With long and accurate HiFi reads, you can characterize the full diversity of the transcriptome:
- See alternative start and end sites
- Characterize splicing events
- Identify allele-specific isoforms
- Profile expression at isoform resolution
- Detect fusion genes
- Predict full-length open read frames
- …and more
“More than 50% of isoforms robustly detected using Iso-Seq could not be readily recapitulated using standard RNA-seq, highlighting the advantage of long-read sequencing for characterizing isoform diversity.”1
– Leung, et al, (2021)
1. Leung, et al. (2021) Full-length transcript sequencing of human and mouse cerebral cortex identifies widespread isoform diversity and alternative splicing. Cell Rep. 37(7):110022.
RNA sequencing with Iso-Seq analysis in action
Article
New isoform phasing technique traces parental-progeny differences in maize
See how Cold Spring Harbor scientists endeavored to ensure that corn progeny could outperform their ‘parents.’
Scalable RNA isoform sequencing using intra-molecular multiplexed cDNAs
For scalable full-length RNA sequencing, we have developed multiplexed arrays sequencing…
Iso-Seq analysis enthusiasts share research wins at virtual social event
Dozens of self-proclaimed Iso-Seq analysis geeks and other curious researchers gathered…
Isoform-level transcriptome reference helps shrimp ward off disease
Shrimp are one of the fastest growing global foods, but also one of the most vulnerable.
EXPLORE
Did you know we have a comprehensive library of articles, reports, papers, and videos related to RNA sequencing with the Iso-Seq method?
SHAPING THE FUTURE
Iso-Seq analysis provides insights into feats of physiology of hibernating bears
Washington State University researchers use Iso-Seq analysis to shed light on the metabolic mystery of brown bears.
Iso-Seq application workflows at a glance
Advantages of RNA sequencing with the
Iso-Seq method
Powered by HiFi reads, the Iso-Seq method uncovers biology that other approaches miss.
Iso-Seq Method delivers full-length transcript isoforms, no assembly required
Unlike short-read RNA-seq methods, Iso-Seq method reads span the whole transcript, capturing even untranslated regions.
- Detect alternative polyadenylation
- Detect differential isoform usages (DIU)
- Obtain isoform-level phasing information
Iso-Seq Data is comprehensive
Characterize whole transcriptome isoform diversity that can reveal complex alternative splicing, fusion events, and transcriptional readthroughs.
Iso-Seq Analysis offers unparalleled insight at the single cell level
Use any single-cell platform to generate full-length cDNA as input for SMRT sequencing and achieve all the same benefits of Iso-Seq on bulk cDNA — complete transcript sequence, isoform-level phasing, and isoform profiling — at single-cell resolution.
TECHNOLOGY DEVELOPMENT SPOTLIGHT
MAS-ISO-Seq method delivers >15× throughput through concatenation
Our technology is constantly evolving. MAS-ISO-Seq data takes advantage of long and accurate HiFi read lengths to increase Iso-Seq throughput by >15-fold through multiplexed arrays of cDNA molecules.
Watch the MAS-ISO-Seq talk at ASHG or read the preprint.
Common questions about PacBio Iso-Seq method and RNA sequencing
Sequel II Systems
Sequel II systems can generate longer reads with greater accuracy and throughput, at a significantly lower cost. Explore our sequencing platforms for full-length cDNA sequencing.