Direct RNA sequencing

Overview:

Direct RNA Sequencing (Nanopore) provides a method for sequencing native RNA molecules without prior reverse transcription or amplification, preserving base modifications and full-length transcript information. Sequencing is performed using Oxford Nanopore long-read platforms which thread single RNA molecules through nanopores to detect electrical signal changes corresponding to nucleotide identity. Requires high-quality RNA, ideally intact. Enables detection of full-length transcripts, splice variants, RNA modifications, polyadenylation sites, and transcript isoform quantification. Unlike short-read RNA-seq, direct RNA sequencing avoids biases introduced by cDNA synthesis or PCR amplification, providing a more faithful representation of transcript structure and modifications. Applicable to RNA extracted from cells, tissues, or biofluids. Supports transcriptome profiling, alternative splicing analysis, RNA modification mapping, viral and microbial RNA sequencing, and isoform-level quantification for functional genomics studies.

Workflow:

RNA extraction, adapter ligation, sequencing.