Nanopore Sequencing: Scientific Principles, Platforms, Protocols, Applications, and Perspectives

Nanopore sequencing has emerged as a game-changing technology that allows scientists to directly read the sequence of DNA and RNA molecules by detecting subtle changes in electrical current as these molecules pass through tiny protein pores. Unlike traditional sequencing methods, it offers the ability to generate ultra-long reads in real time, enabling new insights into complex genomes, RNA transcripts, and chemical modifications-without the need for amplification or extensive laboratory setups. What makes this technology truly remarkable is its portability and versatility, powering everything from field-based pathogen surveillance to largescale population genomics projects. In this review, we explore how nanopore sequencing works, provide a comprehensive overview of the various Oxford Nanopore platforms currently available, and detail practical considerations in sample preparation, data analysis, and interpretation.