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中国生物工程杂志

China Biotechnology
China Biotechnology  2021, Vol. 41 Issue (8): 75-89    DOI: 10.13523/j.cb.2104018
    
Signal Processing for Nanopore Sequencing and Its Application in DNA Data Storage
GE Qi1,ZHANG Peng1,HAN Ming-zhe2,3,YANG Jin-sheng1,ZHANG Da-lu4,*(),CHEN Wei-gang1,3
1 School of Microelectronics, Tianjin University, Tianjin 300072, China
2 School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
3 Frontiers Science Center for Synthetic Biology (MOE), Tianjin University, Tianjin 300072, China
4 China National Center for Biotechnology Development, Beijing 100039, China
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Abstract  

With the continuous update of high-throughput sequencing technologies, the third-generation sequencing technology that can read nucleotide sequences at the single-molecule level has developed rapidly. Nanopore sequencing technology is its representative single-molecule sequencing technology, which realizes base calling by detecting the characteristic changes of electrical current when the DNA single-stranded molecule is passing through a nanopore channel. Compared with the traditional first-generation and the next-generation sequencing (NGS) technologies, the nanopore sequencing of DNA has great advantages in device portability, base acquisition speed and read length, which has attracted much attention. With the continuous development of nanopore sequencing technologies, various signal processing schemes and biological information processing tools for nanopore sequencing have been developed, and base calling and model simulation are two of the key research directions. The fundamental principle and signal processing flow of nanopore sequencing are surveyed, the current challenges are discussed, then the development trend of base calling and nanopore model simulation in recent years are summarized, and the performance of different base calling methods are compared by using real sequencing reads. Then, an integrated simulation platform for the evaluation of signal processing algorithms of nanopore sequencing is developed. Furthermore, with the explosive growth of global data volume, DNA data storage is becoming a promising medium for future massive data storage, and the use of nanopore for sequencing and reading is a very effective method. The application progress of the nanopore sequencing technology for DNA data storage is summarized, and its feasibility is analyzed. The rapid readout method of artificial chromosome data storage based on nanopore sequencing is analyzed, and the application of the simulation of nanopore sequencing reads combined with actual sequencing data in DNA data storage is discussed, which provides a reference for the development of a suitable DNA data storage program.



Key wordsNanopore sequencing      Base calling      Nanopore signal processing      DNA data storage     
Received: 14 April 2021      Published: 31 August 2021
ZTFLH:  Q819  
Corresponding Authors: Da-lu ZHANG     E-mail: zhangdl@cncbd.org.cn
Cite this article:

GE Qi,ZHANG Peng,HAN Ming-zhe,YANG Jin-sheng,ZHANG Da-lu,CHEN Wei-gang. Signal Processing for Nanopore Sequencing and Its Application in DNA Data Storage. China Biotechnology, 2021, 41(8): 75-89.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2104018     OR     https://manu60.magtech.com.cn/biotech/Y2021/V41/I8/75

Fig.1 The workflow and principles of the nanopore sequencing paradigm
碱基识别软件 是否开源 程序语言 输入数据类型 核心计算模型 开发团队
Nanocall C++ 分段事件 隐马尔科夫模型 David等[41]
Nanonet C++ 分段事件 循环神经网络 ONT公司
DeepNano Python 分段事件 循环神经网络 Boza等[42]
BasecRAWller - 原始电流信号 循环神经网络 Stoiber等[43]
Chiron Python 原始电流信号 循环神经网络 Teng等[44]
Albacore Python 原始电流信号 循环神经网络 ONT公司
Guppy - 原始电流信号 循环神经网络 ONT公司
Scrappie C 分段事件或原始电流信号 循环神经网络 ONT公司
Flappie C 原始电流信号 循环神经网络 ONT公司
Table 1 The comparison of base calling software for nanopore sequencing
Fig.2 The performance comparison of base calling software
Fig.3 The memory error model for IDS
仿真软件 程序语言 是否生成仿真读段 是否生成仿真信号 是否模拟噪声特性 开发团队
ReadSim Python Lee等[50]
SiLiCO Python Baker等[51]
NanoSim Python Yang等[52]
DeepSimulator Python Li等[53,54]
NanosigSim Python Chen等[55]
Table 2 The comparison of simulation software for nanopore sequencing
Fig.4 The simulation platform for nanopore sequencing
Fig.5 The verification platform for data readout from the encoded artificial chromosome specific for data storage
Fig.6 The simulation of nanopore sequencing based DNA data storage using real sequencing data for training
Fig.7 The addressing method of simulation data based on nanopore sequencing
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