EUROPIUM-LABELED OLIGONUCLEOTIDE HYBRIDIZATION PROBES - PREPARATION AND PROPERTIES

被引:19
作者
DAHLEN, P
LIUKKONEN, L
KWIATKOWSKI, M
HURSKAINEN, P
IITIA, A
SIITARI, H
YLIKOSKI, J
MUKKALA, VM
LOVGREN, T
机构
[1] WALLAC OY,POB 10,SF-20101 TURKU,FINLAND
[2] UNIV TURKU,DEPT BIOCHEM,SF-20520 TURKU 52,FINLAND
[3] UNIV UPPSALA,CTR BIOMED,DEPT MED GENET,S-75123 UPPSALA,SWEDEN
关键词
D O I
10.1021/bc00027a013
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A chemical method for labeling of oligonucleotide probes with europium chelates is presented. A modified deoxycytidine phosphoramidite is used to introduce multiple reactive amino groups to the oligonucleotide during the synthesis phase. Upon deprotection and purification of the modified oligonucleotide, an isothiocyanate derivative of a stable Eu chelate is reacted with the primary amino groups. The labeling technology enables the coupling of a high number of Eu chelates to a single probe. The melting temperatures and hybridization efficiencies of the oligonucleotides are not significantly altered by the labeling process. However, hybridization kinetics of the oligonucleotides are affected by the introduction of multiple modified deoxycytidine residues. In a solid-phase hybridization assay, Up to 10(7) target molecules can be detected.
引用
收藏
页码:268 / 272
页数:5
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