OHMX_High-throughput OMICS solutions
Direct RNA/cDNA sequencing
Direct RNA/cDNA sequencing

Direct RNA sequencing with the Oxford Nanopore Technologies instruments allows accurate quantification and complete, full-length characterisation of RNA or cDNA. The technology makes it possible to sequence without fragmentation or amplification, removing all sources of bias and thus enabling the identification of epigenetic modifications alongside the nucleotide sequence.

For direct RNA sequencing we request 500ng total RNA input. For direct cDNA sequencing we request a minimum of 250ng total RNA input.

Advantages of direct RNA sequencing

Accurate full length transcripts

Detection of epigenetic modifications

Elimination of PCR bias

Minimal sample handling and rapid library preparation

Workflow

The process starts by performing an initial quality check of the total RNA provided by you. During this quality check, the concentration and integrity of the RNA is determined. We only continue with samples that passed this initial quality check. If for some reason certain samples did not pass this initial check, we will contact you to discuss on how to proceed.

In the next steps, the RNA is captured (Illumina TruSeq RNA Access library prep kit) and transcribed (Illumina TruSeq RNA Access library prep kit / Lexogen QuantSeq 3′ mRNA-Seq Library Prep) to cDNA. From the cDNA, sequencing libraries are constructed, which are again quality checked.

Finally, the libraries are sequenced on one of our Oxford Nanopore Technologies sequencers. The technical quality of the sequencing run is monitored in real time.

Raw sequencing data can be transferred to you via the ohmX.bio server (FTP download).