Nano2NGS: A Framework for Converting Third-Generation Sequencing Data to NGS-Liked Sequencing Data for Hotspot Mutation Detection and Metagenomic Analysis

Nanopore sequencing, also known as single-molecule real-time sequencing, is a third/fourth generation sequencing technology that enables deciphering single DNA/RNA molecules without the polymerase chain reaction. Although third generation sequencing (TGS) has made significant progress in scientific research and clinical practice, its application has been limited compared with next-generation sequencing (NGS) due to specific design principles and data formats, especially in point mutation detection and metagenomic analysis. We therefore developed Nano2NGS as a data analysis framework for hotspot mutation detection and metagenomic analysis based on long reads from nanopore sequencing. Nano2NGS is characterized by applying nanopore sequencing data to NGS data analysis pipelines. Long reads can be converted into short reads using statistical methods and then processed through existing NGS analysis pipelines to obtain more accurate results compared to conventional TGS data analysis algorithms. Nano2NGS serves as a reference and tool for TGS data analysis, such as base recognition and taxonomic classification of metagenomic data based on nanopore sequencing. This framework also promotes higher application scope of TGS in clinical practice.Very important note: Although the data is nanopore sequencing data, the sequencing platform is a new nanopore sequencing platform different from ONT, which is called QNome-9604 platform. Since SRA does not currently have a record of this sequencing platform, the MinION sequencing platform is temporarily selected here. Otherwise, the data cannot be uploaded and made public. If permitted, we will make corrections as soon as possible.