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    Telomeres, the protective caps at the end of our chromosomes, sit at the intersection of aging.
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Oxford Nanopore Technologies Service

cDNA Sequencing

  • A long-read RNA sequencing service for full-length cDNA analysis, isoform discovery, and transcriptome characterization.
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Overview

cDNA Sequencing is a standard Oxford Nanopore Technologies service for long-read sequencing of cDNA reverse transcribed from RNA. The workflow supports full-length transcript, isoform, splice variant, and fusion gene analysis, with UMI-enabled preparation to reduce amplification bias.
  • Full-length transcript and isoform characterization
  • High-throughput multiplexing for cost-efficient sequencing
  • Flexible compatibility with total or enriched RNA inputs
  • UMI-enabled preparation to reduce amplification bias
Full-Length Transcript Characterization
Full-length transcript and isoform analysis
Splice-variant and fusion transcript detection
Quantitative Transcript Profiling
UMI-enabled transcript quantification
Reduced PCR bias for abundance estimation
Research & Translational Applications
Transcriptome characterization and annotation
Biomarker discovery and expression profiling
Technology
Workflow
Performance
Data Outputs

Technology and Compatibility

cDNA Sequencing applies ONT long-read sequencing to cDNA generated from RNA across diverse species and sample types, enabling transcript-level analysis across human, animal, plant, microbial, and non-model species.
  • Compatible with
  • RNA from human, animal, plant, microbial, and non-model organisms
  • Total RNA, poly(A)-enriched RNA, and ribodepleted RNA preparations
  • Research, translational, and transcriptome characterization workflows
  • Component
    Technology
  • Sequencing Chemistry
    Oxford Nanopore cDNA (PCR) Sequencing
  • Preparation
    Standard ONT-compatible cDNA (PCR) preparation
  • Sequencing Platform
    Oxford Nanopore PromethION

Workflow

cDNA Sequencing uses ONT’s long-read cDNA (PCR) sequencing workflow. Renew provides full-service support from RNA extraction through library preparation, sequencing, analysis, and data delivery.
Diagram of cDNA workflow showing full-length RNA processing through adapter ligation, digestion, PCR, and sequencing.
Figure 1. Oxford Nanopore Technologies cDNA PCR Sequencing library preparation workflow (SQK-PCS114).
  • 1. RNA Extraction
    High-quality RNA is isolated from submitted samples.
  • 2. Library Preparation
    RNA is prepared using ONT’s SQK-PCS114 library preparation.
  • 3. Long-Read Sequencing
    cDNA sequencing is performed on the PromethION platform.
  • 4. Data Analysis
    Sequencing data are processed through automated analysis pipelines.
  • 5. Data Delivery
    Results are delivered through the customer portal or AWS S3.

Sequencing Performance

Coverage and yield vary by sample type, input quality, and study goals. Representative performance metrics shown below are based on human tissue samples. cDNA sequencing can also be performed for other species of interest.
Download Spec Sheet
Table comparing Illumina and ONT cDNA sequencing metrics: reads, bases, alignment, length, isoform resolution.
  • Metric
    Typical Performance
  • Samples per Flow Cell
    Up to 24
  • Average Output
    4–6 Gb per sample in a 24-sample run
  • Average Read Length
    1-2kb
  • Available Epigenetic Signals
    Not retained after cDNA conversion

Bioinformatics and Data Outputs

Data outputs are designed to optimize data storage, transfer, and analysis. All outputs are easily accessible through the Client Portal and can be transferred via AWS S3 or other API connections as needed.
Download modal showing files Report, Aligned Bam, and POD5 for specimen 250409 with dates and sizes.
  • Data Outputs
  • QC Report
    Sequencing metrics and run statistics
  • BAM
    Aligned reads
  • POD5
    Raw sequencing data (available for an additional fee)
  • Data Delivery Options
  • Client Web Portal
    Manual download | Mac, Linux, Windows
  • AWS S3 Transfer
    Automated process | AWS to AWS
  • Command Line Script
    Manual download | Mac, Linux

Service Inclusions

Each project is supported by dedicated project management and cross-functional expertise across assay development, bioinformatics, and regulatory strategy. Renew provides proactive communication, transparent project tracking, and technical support from study design through data delivery.
Renew combines platform flexibility, nanopore expertise, and operational excellence to resolve areas
of the genome that standard approaches cannot.
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