Unlocking the Potential of Nanopore Sequencing: Principles, Advances, and Challenges
Rekha Mahato *
ICFRE-Rain Forest Research Institute, Jorhat, Assam-785001, India.
Prem Chand Gyani
ICFRE-Rain Forest Research Institute, Jorhat, Assam-785001, India.
Akash Maity
TMC-Advanced Centre for Treatment, Research and Education in Cancer, Kharghar, Navi Mumbai-410210, India.
Rupesh Kumar
Rubber Board, Williamnagar, Meghalaya-794111, India.
Mitali Mehta
ICFRE-Rain Forest Research Institute, Jorhat, Assam-785001, India.
Pooja Yadav
Bihar Agriculture University, Sabour, Bihar- 813210, India.
Priyasa Gayan
ICFRE-Rain Forest Research Institute, Jorhat, Assam-785001, India.
Rishita Pakhira
University of Padua,TESAF, Legnaro, Padova, 35020, Italy.
*Author to whom correspondence should be addressed.
Abstract
DNA sequencing is nowadays considered as the most effective technique for detecting genetic differences among populations of a species at the molecular level. There are 3 (Three) generations of sequencing available, which can be efficiently utilized to determine genetic variations. Among all the generations of DNA sequencing, nanopore sequencing technology is anticipated to attain all the standard goals of molecular biology. Deamer, Branton, and coworkers in 1961 first demonstrated the DNA translocation using the α-hemolysin nanopore. In 2014, Oxford Nanopore Technologies' (ONT) MinION sequencer, a cutting-edge revolutionary sequencing prototype, flourished in pre-release form. This device is capable of reading mega-bases and has a streaming mode of operation allowing real time read processing. By electrophoretically forcing molecules in solution via a nanoscale pore, a nanopore-based device provides analytical capabilities and aids in single-molecule detection. The ability of nanopore sequencing to enable long reads, real-time outcomes, and flexibility makes it a versatile tool, although challenges like data processing and its accuracy can still be inconsistent. Genomic research has been transformed by recent developments in sequencing technology, such as Minimap2 and GraphMap2. An effective aligner designed for long-read sequencing, Minimap2 yields precise results for PacBio and nanopore data. An improved version GraphMap2, is crucial for complicated genomic analysis because it increases alignment precision for high-error-rate sequences, especially those produced by nanopore technology. By combining long-read and short-read technologies, ONT (Oxford Nanopore Technologies) is a major player in hybrid sequencing, enabling more precise and comprehensive genome assemblies. This review highlights the history and principles of nanopore technology, technological advancements to the time to improve the accuracy, coverage and utility in vast areas. Challenges and Future prospects are also been discussed.
Keywords: Nanopore, MinION, ONT