RNA sequencing
Reveal novel biology that other sequencing methods miss with full-length transcripts using the Iso-Seq™ method
RNA sequencing
With long and accurate HiFi reads, you can characterize the full diversity of the transcriptome — that are tens of kilobases long. That’s enough to capture most transcripts in a single sequencing read, with no downstream bioinformatic assembly required. This became known as the Iso-Seq® method because it enabled full-length isoform RNA sequencing, which can be used to explore a transcriptome in its entirety or in a targeted fashion to examine individual genes.
The Iso-Seq method can be used in RNA sequencing 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
Upcoming webinar
Detecting cancer-related RNA dysregulation with long-read sequencing
Wednesday, May 10, 2023
8.00 AM PDT | 11.00 AM EDT | 4.00 PM BST| 5.00 PM CEST
Full-length, single-cell RNA data provides critical insights into the understanding of cancer transcriptomic features, such as isoforms, fusions, and expressed mutations. However, until recently, cell typing based on gene expression has been challenging with long reads due to insufficient sequencing depth.
Human transcriptome studies
Explore how changes in isoform usages contribute to 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.
“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) Full-length transcript sequencing of human and mouse cerebral cortex identifies widespread isoform diversity and alternative splicing. Cell Rep. 37(7):110022.
Iso-Seq application workflows at a glance
RNA sequencing datasets
| Application | Dataset | Data type | PacBio system |
|---|---|---|---|
| MAS-Seq single cell | Homo sapiens - PBMC 10x Chromium Single Cell 3' libraries | HiFi reads | Sequel II and Revio systems |
| Whole transcriptome | HiFi reads | Sequel II system | |
| Whole transcriptome | HiFi reads | Sequel II system |
RNA sequencing with Iso-Seq analysis in action
Article
ISO-SEQ METHOD DIFFERENTIATES PSEUDOGENES ASSOCIATED WITH DISEASES
Read how researchers at University College London used the Iso-Seq method to accurately map and characterize the parent-pseudogene pair GBA-GBAP1 which are implicated in Gaucher disease and Parkinson’s disease
Scalable RNA isoform sequencing using intra-molecular multiplexed cDNAs
For scalable full-length RNA sequencing, we have developed multiplexed arrays sequencing…
INNOVATIONS ABOUND AT ISO-SEQ SOCIAL CLUB VOL. 3
Read the recap on the latest Iso-Seq research that is pushing the frontiers of cutting-edge science
PROTEIN ISOFORM DISCOVERY ENHANCED BY ISOFORM SEQUENCING
Researchers use Iso-Seq data to predict open reading frames integrated into a proteogenomics workflow to discovery novel protein isoforms to reveal the functional impacts of alternative splicing
EXPLORE
Did you know we have a comprehensive library of articles, reports, papers, and videos related to RNA sequencing with the Iso-Seq method?
SINGLE-CELL RNA SEQUENCING SPOTLIGHT
MAS-SEQ KIT DELIVERS 16x THROUGHPUT FOR SINGLE-CELL ISOFORM SEQUENCING
The MAS-Seq for 10x Single Cell 3’ kit (based on the MAS-Seq method by Al’Khafaji et al.) takes advantage of long and accurate HiFi read lengths to increase single-cell Iso-Seq throughput by 16-fold through concatenated arrays of single-cell cDNA molecules.
Learn more about the MAS-Seq kit now at the single-cell Iso-Seq webpage.
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.
Common questions about PacBio Iso-Seq method and RNA sequencing
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.
FEATURED LONG-READ SEQUENCING SYSTEMS
Achieve highly accurate full-length cDNA sequencing with PacBio long-read sequencers.