Bring certainty to isoform detection
Changes in isoform variants and relative abundancies are an important clue in deciphering how genomic variants drive the phenotypic differences between health and disease. Only long-read sequencing can provide unbiased, direct detection of all the isoforms present in your sample.
Single Molecule, Real-Time (SMRT) Sequencing delivers the long reads needed to capture intact isoform information without assembly or complicated algorithms. Use the Iso-Seq method to:
- Profile whole transcriptomes exhaustively at the isoform-level
- Characterize alternative start and polyadenylation sites as well as exon skipping events
- Detect isoforms as potential biomarkers
- Target genes of interest using synthetic probes with sample multiplexing to query isoforms in a cost-effective manner
 | Sample & Library Preparation |
| Prepare full-length transcripts with flexible sample and library workflows |
 | Sequencing |
| Simplify sample prep workflow and reduce project costs by using the Sequel Systems |
 | Data Analysis |
| Output high-quality, full-length transcript sequences with no assembly required |
Application Brief: Learn more about these best practices for long-read RNA sequencing
Spotlight: Identify novel full-length isoforms in castration-resistant prostate cancer (CRPR)
Scientists used long-read sequencing to identify and quantify full-length androgen receptor (AR) isoforms expressed in CRPC. They identified a truncated isoform, AR-V9, which shows potential to be a more predictive biomarker than AR-V7 of primary resistance to AR-targeted therapies. Explore this research further:
Kohli, M. et al. (2017) Androgen receptor variant AR-V9 is co-expressed with AR-V7 in prostate cancer metastases and predicts abiraterone resistance. Clinical Cancer Research
Selected Resources
- Sheynkman, Gloria M. et al. (2019) ORF Capture-Seq: a versatile method for targeted identification of full-length isoforms bioRxiv
- Watson, Corey T. et al. (2019) Germline murine immunoglobulin IGHV genes in wild-derived and classical inbred strains: a comparison BioRxiv
- Mantere, Tuomo et al. (2019) Long-Read Sequencing Emerging in Medical Genetics Frontiers in genetics
- Dainis, Alexandra et al. (2019) Targeted Long-Read RNA Sequencing Demonstrates Transcriptional Diversity Driven by Splice-Site Variation in MYBPC3. Circulation. Genomic and precision medicine
- Jing, Ying et al. (2019) Hybrid sequencing-based personal full-length transcriptomic analysis implicates proteostatic stress in metastatic ovarian cancer. Oncogene
- Sayed, Mohammed E et al. (2019) NOVA1 directs PTBP1 to hTERT pre-mRNA and promotes telomerase activity in cancer cells. Oncogene
- Lian, Bi et al. (2019) Unveiling novel targets of paclitaxel resistance by single molecule long-read RNA sequencing in breast cancer. Scientific reports
- Deslattes Mays, Anne et al. (2019) Single-Molecule Real-Time (SMRT) Full-Length RNA-Sequencing Reveals Novel and Distinct mRNA Isoforms in Human Bone Marrow Cell Subpopulations. Genes
- Bruijnesteijn, Jesse et al. (2018) Extensive alternative splicing of KIR transcripts. Frontiers in immunology
- Lee, Ming-Hsiang et al. (2018) Somatic APP gene recombination in Alzheimer’s disease and normal neurons. Nature
- Hughes, Timothy et al. (2018) Elevated expression of a minor isoform of ANK3 is a risk factor for bipolar disorder. Translational psychiatry
- Gupta, Ishaan et al. (2018) Single-cell isoform RNA sequencing characterizes isoforms in thousands of cerebellar cells. Nature biotechnology
- Pike, Alexandra M et al. (2018) TIN2 functions with TPP1/POT1 to stimulate telomerase processivity BioRxiv
- Dougherty, Max L et al. (2018) Transcriptional fates of human-specific segmental duplications in brain. Genome research
- Pollard, Martin O et al. (2018) Long reads: their purpose and place. Human molecular genetics
- Nattestad, Maria et al. (2018) Complex rearrangements and oncogene amplifications revealed by long-read DNA and RNA sequencing of a breast cancer cell line. Genome research
- Kronenberg, Zev N et al. (2018) High-resolution comparative analysis of great ape genomes. Science
- Cantsilieris, Stuart et al. (2018) Recurrent structural variation, clustered sites of selection, and disease risk for the complement factor H (CFH) gene family. Proceedings of the National Academy of Sciences of the United States of America
- Anvar, Seyed Yahya et al. (2018) Full-length mRNA sequencing uncovers a widespread coupling between transcription initiation and mRNA processing. Genome biology
- Bruijnesteijn, Jesse et al. (2018) Human and rhesus macaque KIR haplotypes defined by their transcriptomes. Journal of immunology
- Ardui, Simon et al. (2018) Single molecule real-time (SMRT) sequencing comes of age: applications and utilities for medical diagnostics. Nucleic acids research
- Asur, Rajalakshmi S et al. (2018) Somatic mosaicism of an intragenic FANCB duplication in both fibroblast and peripheral blood cells observed in a Fanconi anemia patient leads to milder phenotype. Molecular genetics & genomic medicine
- Tseng, Elizabeth et al. (2019) The landscape of SNCA transcripts across synucleinopathies: New insights from long reads sequencing analysis BioRxiv
- Gao, Shan et al. (2018) Two novel lncRNAs discovered in human mitochondrial DNA using PacBio full-length transcriptome data. Mitochondrion
- Poster: Tseng, Elizabeth et al. (2019) Full-length transcriptome sequencing of melanoma cell line complements long-read assessment of genomic rearrangements
- Poster: Liqing, Tian et al. (2018) Allelic specificity of immunoglobulin heavy chain (IGH@) translocation in B-cell acute lymphoblastic leukemia (B-ALL) unveiled by long-read sequencing
- Poster: Ashby, M. et al. (2018) Scalability and reliability improvements to the Iso-Seq analysis pipeline enables higher throughput sequencing of full-length cancer transcripts
- Poster: Ho, Yeung et al. (2018) The role of androgen receptor variant AR-V9 in prostate cancer
- Poster: Ekholm, J. et al. (2017) Screening for causative structural variants in neurological disorders using long-read sequencing
- Poster: Ashby, M. et al. (2017) Simplified sequencing of full-length isoforms in cancer on the PacBio Sequel platform
- Poster: Clark, Tyson A. et al. (2017) SMRT Sequencing of full-length androgen receptor isoforms in prostate cancer reveals previously hidden drug resistant variants
- Poster: Kujawa, Steve et al. (2017) A method for the identification of variants in Alzheimer’s disease candidate genes and transcripts using hybridization capture combined with long-read sequencing
- Poster: Ashby, Meredith et al. (2017) Using the PacBio IsoSeq method to search for novel colorectal cancer biomarkers
- Badgett, Marty (2019) AGBT Presentation: The Sequel II System – the next evolution of SMRT Sequencing
- Korlach, Jonas and Corney, Dave (2018) Webinar: A new paradigm in DNA sequencing – Highly accurate single-molecule long reads
- (2018) Tutorial: Iso-Seq analysis application [SMRT Link v6.0.0]
- Tseng, Elizabeth (2018) ASHG PacBio Workshop: The Iso-Seq method for discovering alternative splicing in human diseases
- Pillai, Nisha (2018) Webinar: Survey of transcriptome diversity using Iso-Seq analysis
- Ashby, Meredith (2017) Webinar: Chasing alternative splicing in cancer: Simplified full-length isoform sequencing
- Auvignen, Petra (2017) AGBT Virtual Poster: Interspecies interation amoung meat spoilage-related lactic acid bacteria
- Ashby, Meredith (2017) AGBT Virtual Poster: Using the PacBio Iso-Seq method to search for novel colorectal cancer biomarkers
- Application Brief: Long-read RNA sequencing – Best Practices (2019)
- PacBio Certified Service Providers (2019)
- SMRT Sequencing Brochure: Delivering highly accurate long reads to drive discovery in life science (2019)
- Cancer Brochure: Sequence cancer variants with confidence (2019)
- SMRT Analysis Brochure: Gain a deeper understanding of your sequencing data (2018)
- Case Study: SMRT sequencing brings clarity to HIV vaccine and transplant research at the Wisconsin national primate research center (2017)