Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes
Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes
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DOI：
10.1093/nar/gnf121
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发表时间：
2002-11-01
影响因子：
14.9
通讯作者：
Tyagi, S
中科院分区：
文献类型：
--
作者：
Marras, SAE;Kramer, FR;Tyagi, S
在设计用于均相杂交测定的寡核苷酸探针中的一个重要考虑因素是用于标记探针的荧光团和猝灭剂之间的能量转移效率。我们已经确定了大量的常用的荧光团和猝灭剂的组合的能量转移的效率。我们还测量了核苷酸对每个荧光团的荧光的猝灭效应。测量了共振能量转移和静态猝灭模式的猝灭效率。我们发现，除了它们的光化学特性，荧光团和猝灭剂相互结合的趋势对猝灭效率有很强的影响。这些测量的可用性应有助于设计含有相互作用的荧光团和猝灭剂的寡核苷酸探针，包括竞争性杂交探针、邻近探针、TaqMan探针和分子信标。
An important consideration in the design of oligonucleotide probes for homogeneous hybridization assays is the efficiency of energy transfer between the fluorophore and quencher used to label the probes. We have determined the efficiency of energy transfer for a large number of combinations of commonly used fluorophores and quenchers. We have also measured the quenching effect of nucleotides on the fluorescence of each fluorophore. Quenching efficiencies were measured for both the resonance energy transfer and the static modes of quenching. We found that, in addition to their photochemical characteristics, the tendency of the fluorophore and the quencher to bind to each other has a strong influence on quenching efficiency. The availability of these measurements should facilitate the design of oligonucleotide probes that contain interactive fluorophores and quenchers, including competitive hybridization probes, adjacent probes, TaqMan probes and molecular beacons.