In this study cDNA of HEK293 and HeLa cells was sequenced in order to provide test data for a nanopore sequencing real-time transcriptional analysis pipeline. (NanopoReaTA)

Oxford Nanopore Technologies' (ONT) sequencing platform (Nanopore-Seq) offers the great opportunity to perform real-time analysis during sequencing. This feature allows for early insights into experimental data and accelerates a potential decision-making process for further analysis, which can be particularly relevant in the clinical context. Despite the development of novel tools for real-time analysis of Nanopore-Seq data from DNA, there is currently no application specialized for transcriptional analysis designed for scientists or physicians of limited bioinformatics knowledge. Here we introduce NanopoReaTA, a user-friendly real-time analysis toolbox for RNA sequencing data from ONT. Sequencing results from a running or finished experiment are processed through an R Shiny-based graphical user interface (GUI) with an integrated Nextflow pipeline for whole transcriptome and/or gene-specific analyses. NanopoReaTA provides visual snapshots of a sequencing run in progress, enabling rapid decision making that could potentially be applied to clinical cases. The provided dataset includes ONT based direct cDNA sequencing data of HEK293 and HeLa cells to prove the functionality of NanopoReaTA.