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使用荧光DNA探针检测成熟RNA而非前体RNA。

Detection of the mature, but not precursor, RNA using a fluorescent DNA probe.

作者信息

Paiboonskuwong Kittiphong, Kato Yoshio

机构信息

Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba Science City 305-8562, Japan.

出版信息

Nucleic Acids Symp Ser (Oxf). 2006(50):327-8. doi: 10.1093/nass/nrl163.

Abstract

Fluorescently labelled oligonucleotide probes have been widely used in biotechnology and diagnostic field. We improved the molecular beacon probe to detect the mature, but not precursor, RNA selectively. Based on the principle that a kind of fluorophore can be quenched by adjacent guanine base, therefore, the noise fluorescence caused by the false targets can be avoided. As an example, we designed the probe for the selective detection of a mature microRNA (miRNA). We demonstrate that the probe detects mature miRNA with high specificity. Since the probe itself does not fluoresce but becomes fluorescent upon hybridization to the mature target RNA, it provides a quick and selective way for detection of various forms of RNAs.

摘要

荧光标记的寡核苷酸探针已广泛应用于生物技术和诊断领域。我们改进了分子信标探针,以选择性地检测成熟RNA而非前体RNA。基于一种荧光团可被相邻鸟嘌呤碱基淬灭的原理,因此,可以避免由假靶标引起的噪声荧光。例如,我们设计了用于选择性检测成熟微小RNA(miRNA)的探针。我们证明该探针能以高特异性检测成熟miRNA。由于该探针本身不发荧光,但在与成熟靶标RNA杂交后会发出荧光,它为检测各种形式的RNA提供了一种快速且选择性的方法。

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