单分子实时测序技术的原理与应用

doi:10.16288/j.yczz.14-323

遗传 ›› 2015, Vol. 37 ›› Issue (3): 259-268.doi: 10.16288/j.yczz.14-323

单分子实时测序技术的原理与应用

柳延虎¹,王璐^1,2,于黎^1,2

1. 云南大学,云南省生物资源保护与利用重点实验室,昆明 650091; 2. 云南大学,云南省高校动物遗传多样性与进化重点实验室,昆明 650091

收稿日期:2014-09-25 修回日期:2014-12-01 出版日期:2015-03-20 发布日期:2015-02-10
通讯作者: 于黎,研究员,博士生导师,研究方向：动物遗传与进化。E-mail: yuli-1220@163.com
王璐,助理研究员,研究方向：基因组学。E-mail: wanglu@ynu.edu.cn E-mail:liuyanhu005@163.com
作者简介:柳延虎,博士研究生,研究方向：基因组学。E-mail: liuyanhu005@163.com
基金资助:
国家自然科学基金重大研究计划项目(编号：91131904)和中国科学院昆明动物研究所遗传资源与进化国家重点实验室开放课题(编号：GREKF14-04)资助

The principle and application of the single-molecule real-time sequencing technology

Yanhu Liu¹, Lu Wang^{1, 2}, Li Yu^{1, 2}

1. Laboratory for Conservation and Utilization of Bio-resources, Yunnan University, Kunming 650091,China;
2. Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming 650091, China

Received:2014-09-25 Revised:2014-12-01 Online:2015-03-20 Published:2015-02-10

摘要/Abstract

摘要： 单分子DNA测序技术是近10年发展起来的新一代测序技术,也称为第三代测序技术,包括单分子实时测序、真正单分子测序、单分子纳米孔测序等技术。文章介绍了单分子实时(Single-molecule real-time,SMRT)测序技术的基本原理、性能以及应用。与Sanger测序法和下一代测序技术相比,SMRT测序具有超长读长、测序周期短、无需模板扩增和直接检测表观修饰位点等特点,为研究人员提供了新选择。同时,SMRT测序的低准确率备受争议(约85%),其中约93%的错误是插入缺失,因此,其数据应用于基因组组装前需先对数据进行纠错处理。目前,SMRT测序在小型基因组从头测序和完整组装中已有良好应用,并且已经或将在表观遗传学、转录组学、大型基因组组装等领域发挥其优势,促进基因组学的研究。

关键词: 单分子测序, PacBio, SMRT测序

Abstract: Last decade witnessed the explosive development of the third-generation sequencing strategy, including single-molecule real-time sequencing (SMRT), true single-molecule sequencing (tSMSTM) and the single-molecule nanopore DNA sequencing. In this review, we summarize the principle, performance and application of the SMRT sequencing technology. Compared with the traditional Sanger method and the next-generation sequencing (NGS) technologies, the SMRT approach has several advantages, including long read length, high speed, PCR-free and the capability of direct detection of epigenetic modiﬁcations. However, the disadvantage of its low accuracy, most of which resulted from insertions and deletions, is also notable. So, the raw sequence data need to be corrected before assembly. Up to now, the SMRT is a good fit for applications in the de novo genomic sequencing and the high-quality assemblies of small genomes. In the future, it is expected to play an important role in epigenetics, transcriptomic sequencing, and assemblies of large genomes.

Key words: single molecule sequencing, PacBio, SMRT sequencing

柳延虎, 王璐, 于黎. 单分子实时测序技术的原理与应用[J]. 遗传, 2015, 37(3): 259-268.

Yanhu Liu, Lu Wang, Li Yu. The principle and application of the single-molecule real-time sequencing technology[J]. HEREDITAS(Beijing), 2015, 37(3): 259-268.

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单分子实时测序技术的原理与应用

The principle and application of the single-molecule real-time sequencing technology

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