A real-time assay for DNA sticky-end pairing using molecular beacons
A real-time assay for DNA sticky-end pairing using molecular beacons
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DOI：
10.1016/s0003-2697(02)00375-5
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发表时间：
2003-01-15
影响因子：
2.9
通讯作者：
Tan, WH
中科院分区：
文献类型：
--
作者：
Li, JWJ;Tan, WH
DNA粘端配对(SEP)1在细胞过程和各种生物技术中扮演着重要的角色[1-4]，人们已经用不同的方法进行了研究，包括超速离心法、凝胶电泳法、电子显微镜和原子力显微镜[5-8]。然而，这些方法需要产物分离、DNA连接和表面沉积，这些都会改变SEP的反应平衡。此外，这些方法是不连续的、耗时和费力的。因此，对原位、连续、方便、实时的体感诱发电位分析已有明显的需求。在这里，我们介绍了一种基于分子信标(MB)的有效实时分析方法，用于SEP观察。MB是一种新开发的单链DNA探针，在基因检测和蛋白质定量方面有着广泛的应用[9-13]。MBS的诸多优点之一是“无需分离即可检测”和在均相溶液中的超灵敏分析[11]，这非常适合于实时SEP研究。
DNA sticky-end pairing (SEP) 1 plays an important role in cellular processes and various biotechnologies [1–4] and has been studied with different methods including ultracentrifuge, gel electrophoresis, electron microscopy, and atomic force microscopy [5–8]. However, these methods need product separation, DNA ligation, and surface deposition, all of which tend to shift the reaction equilibrium of SEP. Moreover, these methods are discontinuous, time consuming, and laborious. Therefore, there has been a clear need for in situ, continuous and convenient, real-time SEP assays. Here we introduce an effective real-time assay, based on the molecular beacon (MB), for SEP observation. The MB is a newly developed ssDNA probe and has found wide applications from gene detection to protein quantitation [9–13]. Among the many advantages of MBs are ‘‘detection without separation’’and ultrasensitive analysis in homogeneous solution [11], which fits well for real-time SEP study.