Human RNA sequencing
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.
Profile transcriptome complexity without assembly
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
Workflow: from RNA to full-length transcripts
Sample + library preparation
Prepare full-length transcripts with flexible sample and library workflows
- Use the SMRTbell express template prep kit 3.0 to prepare libraries in one day
- Follow the Iso-Seq library protocol using SMRTbell prep kit 3.0
Sequencing
Simplify sample prep workflow and reduce project costs by using the Sequel II and IIe systems. Generate up to 4 million full-length reads per SMRT Cell 8M*
* Read lengths, number of reads, data per SMRT Cell, and other sequencing performance results vary based on sample quality/type and insert size, among other factors.
Data analysis
Output high-quality, full-length transcript sequences; no assembly required.
- Generate highly accurate long reads (HiFi reads) to characterize whole transcriptomes
- Dataset: Universal Human Reference RNA (UHRR)
- Dataset: Alzheimer Brain
- Use the Iso-Seq analysis in SMRT Link to output high-quality, full-length transcript FASTA sequences, with no assembly required, to characterize transcripts and splice variants
- Run Iso-Seq analysis with or without a reference genome. Follow up with community tools for different analyses such as SQANTI3 (transcript classification), tappAS (differential expression analysis), and TAMA (annotation).
Whitepaper
Bulk and single-cell isoform sequencing for human disease research
In this whitepaper, we discuss the features of bulk and single-cell isoform sequencing with the Iso-Seq method and their applications in human genomics.
Spotlight
IDENTIFY NOVEL FULL-LENGTH ISOFORMS IN BREAST CANCER CLINICAL AND CELL LINE SAMPLES LINKED TO SURVIVAL OUTCOMES
Scientists used the Iso-Seq method to identify novel isoforms uniquely associated with breast cancer subtypes and survival outcomes.
Veiga et al., “A comprehensive long-read isoform analysis platform and sequencing resource for breast cancer“, Science Advances (2022)
