Long-Read Nanopore Sequencing of RPGR ORF15 is Enhanced Following DNase I Treatment of MinION Flow Cells.
Samar YahyaChristopher Mark WatsonIan M CarrMartin McKibbinLaura A CrinnionMorag TaylorHope BoninTracy FletcherMohammed E El-AsragManir AliCarmel ToomesChris F InglehearnPublished in: Molecular diagnosis & therapy (2023)
We report the novel finding that long-read nanopore sequencing can read through RPGR-ORF15, a DNA sequence not captured by short-read next-generation sequencing (NGS), but with a more reduced yield. Use of a flow cell wash kit containing DNase I unblocks the pores, allowing reloading of further library aliquots over a 72-h period, increasing yield. The workflow we describe provides a novel solution to the need for a rapid, robust, scalable, cost-effective ORF15 screening protocol.