Genomics and Applied Biology 2024, Vol.15, No.1, 22-26 http://bioscipublisher.com/index.php/gab 23 1 Overview of Portable Nanopore Sequencing Technology 1.1 Principle of the nanopore technology Portable nanopore sequencing technology is a revolutionary method of genome sequencing that utilizes tiny channels (nanopores) to detect the DNA or RNA sequences of individual molecules. Its working principle is based on guiding single-stranded DNA or RNA molecules through a nanopore embedded in a membrane made of a resistive material. As the molecule passes through the nanopore, it causes changes in the electrical current. These changes are specifically associated with the nucleotide sequence passing through the pore, thereby allowing real-time sequence identification of the molecules passing through the nanopore (Cummings et al., 2017). During the sequencing process, a voltage is applied across the membrane containing the nanopore, prompting the charged molecules to pass through the pore. As the molecules pass through the nanopore, the changes in current caused by the nucleotides are detected and recorded. By analyzing the patterns of these electrical signals, the nucleotide sequence of the molecule can be determined (Figure 1). Figure 1 Schematic of DNA sequencing using a nanopore. DNA passing through a nanopore membrane with electrical signal generation and base identification (Cummings et al., 2017) 1.2 Technical features The main advantages of portable nanopore sequencing technology include its compact size, portability, independence from biochemical reagents, and the ability to perform readings using physical methods directly. This technology is less complex, cost-effective, and capable of sequencing purified genomic DNA, PCR amplicons, cDNA samples, or RNA in real-time and rapidly (Cummings et al., 2017). Additionally, this technology also enables single-molecule sequencing without PCR (Chen et al., 2023). 1.3 Technology development Since the first observation of nucleic acids translocating through nanopores in the 1990s, nanopore sequencing technology has evolved from laboratory experiments to commercial tools. Although the current technology has not yet achieved single-nucleotide resolution sequencing of deoxyribonucleic acid, there have been multiple reports using α-hemolysin protein nanopores for basic DNA analysis, as well as the manufacture of various synthetic nanopores. The commercialization of these technologies has begun, turning nanopore sequencing devices into an inexpensive, fast genomic tool that not only allows for reading lengths exceeding 100 kb but also features portability, low cost, and speed (Rhee and Burns, 2006; Bayley, 2015). Portable nanopore sequencing technology has shown its potential in multiple fields, including microbial diversity research, disease diagnostics, drug target discovery, species conservation, SARS-CoV-2 detection, and applications in microgravity environments (Benítez-Páez et al., 2015; McIntyre et al., 2015; Chen et al., 2023). Furthermore, this technology has also demonstrated potential in forensic analysis, potentially becoming a viable solution for small to medium-sized forensic laboratories (Hall et al., 2020). As nanopore sequencing technology continues to improve and application tools are integrated, its incorporation into laboratory or remote field workflows will further simplify the sequencing process (Pomerantz et al., 2018; Hall et al., 2020).
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