Fig. 1: The development process of nanopore sequencing and its technical advantages compared with other high-throughput sequencing platforms.

A The major events in the development of nanopore sequencing. It has taken 25 years from the concept of nanopore sequencing to the advent of the first nanopore sequencer ONT MinION. During the development of nanopore sequencing, four major technical points have been overcome, respectively making DNA molecular through the nanopores, distinguishing of purines and pyrimidines, recognizing a single base in an individual DNA strand, and controlling the rate of DNA passage through the nanopore. Since 2014, nanopore sequencing technology has been in a rapid development stage, and the throughput and accuracy of nanopore sequencing have greatly improved. In 2020, QitanTech, a Chinese company, announced China’s first nanopore sequencing platform, QNome-9606. After four years, Chinese MGI released the nanopore sequencing platform CycloneSEQ. B The development of the accuracy of nanopore sequencing. In the early stages, to improve the sequencing accuracy, the library preparation of nanopore sequencing usually uses a 2 d strategy, which means both strands of a dsDNA molecule to be sequenced. With the improvement of sequencing protein, the accuracy of single strand reads gradually catches up to the consensus sequences of the 2 d strategy. Of note, the accuracy of QitanTech nanopore sequencing has a significant improvement within the past two years. C Comparison of the advantages of nanopore sequencing relative to other sequencing platforms. Nanopore sequencing have many technological advantages including flexible, producing ultra-long sequencing reads and real-time sequencing.