Ergen Erhan, Weber Markus, Jacob Josemon, Herrmann Andreas, Müllen Klaus
Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
Chemistry. 2006 May 3;12(14):3707-13. doi: 10.1002/chem.200501526.
Single-nucleotide polymorphisms (SNPs) are the most common form of DNA sequence variation. There is a strong interest from both academy and industry to develop rapid, sensitive and cost effective methods for SNP detection. Here we report a novel structural concept for DNA detection based on fluorescence dequenching upon hybridization. The so-called "twin probe" consists of a central fluorene derivative as fluorophore to which two identical oligonucleotides are covalently attached. This probe architecture is applied in homogeneous hybridization assays with subsequent fluorescence spectroscopic analysis. The bioorganic hybrid structure is well suited for sequence specific DNA detection and even SNPs are identified with high efficiency. Additionally, the photophysical properties of the twin probe were investigated. The covalent attachment of two single stranded oligonucleotides leads to strong quenching of the central fluorescence dye induced by the nucleobases. The twin probe is characterized by supramolecular aggregate formation accompanied by red-shifted emission and broad fluorescence spectra.
单核苷酸多态性（SNP）是DNA序列变异最常见的形式。学术界和工业界都对开发快速、灵敏且经济高效的SNP检测方法有着浓厚兴趣。在此，我们报告一种基于杂交时荧光去猝灭的新型DNA检测结构概念。所谓的“双探针”由一个作为荧光团的中心芴衍生物组成，两个相同的寡核苷酸共价连接于其上。该探针结构应用于均相杂交分析及随后的荧光光谱分析。这种生物有机杂化结构非常适合序列特异性DNA检测，甚至能高效识别SNP。此外，还研究了双探针的光物理性质。两个单链寡核苷酸的共价连接导致核碱基对中心荧光染料产生强烈猝灭。双探针的特征是形成超分子聚集体，伴随发射峰红移和宽荧光光谱。
