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单链长 DNA 探针的量子点标记

One-to-one quantum dot-labeled single long DNA probes.

机构信息

State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, PR China.

出版信息

Biomaterials. 2011 Aug;32(23):5471-7. doi: 10.1016/j.biomaterials.2011.04.013. Epub 2011 May 5.

Abstract

Quantum dots (QDs) have been received most attention due to their unique properties. Constructing QDs conjugated with certain number of biomolecules is considered as one of the most important research goals in nanobiotechnology. In this study, we report polymerase chain reaction (PCR) amplification of primer oligonucleotides bound to QDs, termed as QD-based PCR. Characterization of QD-based PCR products by gel electrophoresis and atomic force microscopy showed that QD-labeled long DNA strands were synthesized and only a single long DNA strand was conjugated with a QD. The QD-based PCR products still kept fluorescence properties. Moreover, the one-to-one QD-labeled long DNA conjugates as probes could detect a single-copy gene on maize chromosomes by fluorescence in situ hybridization. Labeling a single QD to a single long DNA will make detection of small single-copy DNA fragments, quantitative detection and single molecule imaging come true by nanotechnology, and it will promote medical diagnosis and basic biological research as well as nano-material fabrication.

摘要

量子点(QDs)由于其独特的性质而受到了极大的关注。构建与一定数量的生物分子偶联的量子点被认为是纳米生物技术中最重要的研究目标之一。在这项研究中,我们报告了聚合酶链反应(PCR)扩增与量子点结合的引物寡核苷酸,称为基于量子点的 PCR。通过凝胶电泳和原子力显微镜对基于 QD 的 PCR 产物的表征表明,合成了带有 QD 标记的长 DNA 链,并且只有一条长 DNA 链与 QD 偶联。QD 标记的长 DNA 偶联物作为探针仍然具有荧光性质。此外,通过荧光原位杂交,QD 标记的单链长 DNA 探针可检测到玉米染色体上的单拷贝基因。将单个 QD 标记到单个长 DNA 上,可以通过纳米技术实现对小的单拷贝 DNA 片段的检测、定量检测和单分子成像,从而促进医学诊断和基础生物学研究以及纳米材料的制造。

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