Direct RNA Sequencing

Overview

Direct RNA Sequencing (DRS) enables the sequencing of native RNA molecules without PCR amplification, preserving transcript structure and RNA modifications that are often lost during conventional cDNA-based RNA sequencing. Using Oxford Nanopore Technologies' PromethION platform, this approach provides full-length transcript information, allowing accurate characterization of isoforms, alternative splicing events, and transcript diversity. This service is ideal for studies focused on transcriptome profiling, RNA modification analysis, long non-coding RNAs, viral transcriptomics, and isoform discovery. Direct sequencing of native RNA minimizes biases introduced during reverse transcription and amplification, providing a more comprehensive view of RNA biology.

Highlights

• Direct sequencing of native RNA molecules without PCR amplification.

• Preserves RNA modifications and transcript structure for epi-transcriptomic analysis (for example: m⁶A).

• Full-length transcript sequencing enables accurate isoform and alternative splicing characterization.

• Compatible with poly(A)-enriched RNA or total RNA samples.

• Supports transcript discovery, RNA modification analysis, poly(A) tail assessment, and gene expression profiling.

• Multiplexing available up to 24 samples

Sample submission requirements

• Please provide samples in nuclease-free water.

• Concentration must be measured by Qubit.

• The core has the right to determine whether submitted samples are qualified for the experiment.

• Please check the core ([email protected] or 713-500-7933) for more information.

Service workflow

Multiplexing service workflow

Data analysis

Downstream analysis can be performed using EPI2ME workflows designed for direct RNA data or custom bioinformatic pipelines.

Useful documents

• Original protocol:
• Direct RNA sequencing (SQK-RNA004) (PDF).
• https://nanoporetech.com/document/direct-rna-barcoding-sqk-drb004-24