Products & Services


Nanopore sequencing offers advantages in all areas of research. Our offering includes DNA sequencing, as well as RNA and gene expression analysis and future technology for analysing proteins.

Learn about applications
View all Applications
News Explore

Assessing the utility of long-read nanopore sequencing for rapid and efficient characterisation of mobile element insertions


Date: 28th September 2020 | Source: Laboratory Investigation

Authors: Christopher M. Watson, Laura A. Crinnion, Helen Lindsay, Rowena Mitchell, Nick Camm, Rachel Robinson, Caroline Joyce, George A. Tanteles, Domhnall J. O’ Halloran, Sergio D. J. Pena, Ian M. Carr, David T. Bonthron.

Short-read next generation sequencing (NGS) has become the predominant first-line technique used to diagnose patients with rare genetic conditions. Inherent limitations of short-read technology, notably for the detection and characterization of complex insertion-containing variants, are offset by the ability to concurrently screen many disease genes. “Third-generation” long-read sequencers are increasingly being deployed as an orthogonal adjunct technology, but their full potential for molecular genetic diagnosis has yet to be exploited.

Here, we describe three diagnostic cases in which pathogenic mobile element insertions were refractory to characterization by short-read sequencing.

To validate the accuracy of the long-read technology, we first used Sanger sequencing to confirm the integration sites and derive curated benchmark sequences of the variant-containing alleles. Long-read nanopore sequencing was then performed on locus-specific amplicons.

Pairwise comparison between these data and the previously determined benchmark alleles revealed 100% identity of the variant-containing sequences. We demonstrate a number of technical advantages over existing wet-laboratory approaches, including in silico size selection of a mixed pool of amplification products, and the relative ease with which an automated informatics workflow can be established.

Our findings add to a growing body of literature describing the diagnostic utility of long-read sequencing.

Read the full text Read the structural variation white paper Find out more about Flongle

Recommended for you

Open a chat to talk to our sales team