SOLID-PHASE SYNTHESIS OF CHELATE-LABELED OLIGONUCLEOTIDES - APPLICATION IN TRIPLE-COLOR LIGASE-MEDIATED GENE ANALYSIS

被引:28
作者
KWIATKOWSKI, M [1 ]
SAMIOTAKI, M [1 ]
LAMMINMAKI, U [1 ]
MUKKALA, VM [1 ]
LANDEGREN, U [1 ]
机构
[1] UNIV TURKU,DEPT CHEM,TURKU,FINLAND
关键词
D O I
10.1093/nar/22.13.2604
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Oligonucleotides labelled with detectable groups are essential tools in gene detection. We describe here the synthesis of pyrimidine deoxynucleotide-building blocks, modified at their C-5 position with a protected form of a strongly chelating agent. These reagents can be used to introduce multiple metal ions into oligodeoxynucleotides during standard oligonucleotide synthesis. The chelating functions form strongly fluorescent complexes with europium ions, characterized by a wide separation between the excitation and emission spectra. Moreover, the long decay time of the fluorescence permits sensitive time-resolved fluorescence measurements. The chelates also have the stability required to function in triple-color assays involving europium, samarium, and terbium ions. We demonstrate the application of these reagents for ligase-based gene analysis reactions.
引用
收藏
页码:2604 / 2611
页数:8
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