摘要
焦测序技术是一种实时DNA测序技术。它在DNA聚合酶、三磷酸腺苷硫酸化酶、荧光素酶和三磷酸腺苷双磷酸酶4种酶的协同作用下,将焦磷酸转化为等量的荧光信号,通过荧光信号的高低实时检测待测序列,操作简便,可实现高通量、自动化测定,检测不需要电泳,不需要对样品标记和染色,结果准确可靠重复性好。本文综述了焦测序技术的基本原理、历史及其在测序模板制备技术、反应体系和检测仪器三个方面的最新进展,并重点介绍制备单链的Late-PCR技术、高灵敏度反应体系的获得以及454公司超高通量测序技术,并对焦测序技术的发展做了展望。
Pyrosequencing is a real-time DNA sequencing technology based on sequencing-by-synthesis.The technique based on a cascade of 4-enzyme reactions to proportionally convert pyrophosphate （PPi） into fluorescent signals.The heights of the peak signals stand for the number of nucleotide,so the unknown sequence from the pyrogram could be precisely read out.Pyrosequencing is characterized by easy handling,high throughput and automation.The advances on pyrosequencing technology were reviewed in this article,including the general principles,the history,the template preparation methods,the developments of pyrosequencing reaction systems,the instruments,the potential applications and developments.
出处
《现代生物医学进展》
CAS
2008年第8期1573-1576,1598,共5页
Progress in Modern Biomedicine
