Nanopore sequencing technology has seen significant advancements, particularly in Oxford Nanopore Technologies' (ONT) MinION device, which has revolutionized the field of long-read sequencing. This technology relies on a nanoscale protein pore embedded in an electrically resistant polymer membrane, allowing the detection of ionic currents as DNA or RNA molecules pass through. The accuracy, read length, and throughput of ONT data have improved over time, driven by advancements in nanopore design, motor proteins, and bioinformatics methods. The review covers the development of nanopore sequencing technology, including the engineering of nanopores and motor proteins, and the improvements in data quality through various strategies. Bioinformatics methods, such as base calling, error correction, and alignment, have been crucial for enhancing the utility of ONT data. The technology is now applied in a wide range of applications, including genome assembly, transcriptome analysis, epigenome and epitranscriptome studies, and clinical diagnostics. Key applications include closing gaps in reference genomes, identifying large structural variations, characterizing full-length transcriptomes, and detecting epigenetic marks. ONT sequencing has also been used in cancer research, infectious disease surveillance, and genetic disease diagnosis. The portable and affordable nature of ONT devices has enabled real-time genomic surveillance of emerging infectious diseases, such as Ebola and Zika virus outbreaks. Despite these advancements, challenges remain, including improving data quality, reducing errors, and developing more efficient and accurate bioinformatics tools. The review concludes by discussing future directions for overcoming these limitations and leveraging the unique strengths of nanopore sequencing.Nanopore sequencing technology has seen significant advancements, particularly in Oxford Nanopore Technologies' (ONT) MinION device, which has revolutionized the field of long-read sequencing. This technology relies on a nanoscale protein pore embedded in an electrically resistant polymer membrane, allowing the detection of ionic currents as DNA or RNA molecules pass through. The accuracy, read length, and throughput of ONT data have improved over time, driven by advancements in nanopore design, motor proteins, and bioinformatics methods. The review covers the development of nanopore sequencing technology, including the engineering of nanopores and motor proteins, and the improvements in data quality through various strategies. Bioinformatics methods, such as base calling, error correction, and alignment, have been crucial for enhancing the utility of ONT data. The technology is now applied in a wide range of applications, including genome assembly, transcriptome analysis, epigenome and epitranscriptome studies, and clinical diagnostics. Key applications include closing gaps in reference genomes, identifying large structural variations, characterizing full-length transcriptomes, and detecting epigenetic marks. ONT sequencing has also been used in cancer research, infectious disease surveillance, and genetic disease diagnosis. The portable and affordable nature of ONT devices has enabled real-time genomic surveillance of emerging infectious diseases, such as Ebola and Zika virus outbreaks. Despite these advancements, challenges remain, including improving data quality, reducing errors, and developing more efficient and accurate bioinformatics tools. The review concludes by discussing future directions for overcoming these limitations and leveraging the unique strengths of nanopore sequencing.