CYANINE DYE DUTP ANALOGS FOR ENZYMATIC LABELING OF DNA PROBES
CYANINE DYE DUTP ANALOGS FOR ENZYMATIC LABELING OF DNA PROBES
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DOI：
10.1093/nar/22.15.3226
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发表时间：
1994-08-11
影响因子：
14.9
通讯作者：
WAGGONER, AS
中科院分区：
文献类型：
--
作者：
YU, H;CHAO, J;WAGGONER, AS
荧光原位杂交(FISH)已成为各种研究领域和临床诊断中不可缺少的工具。许多应用需要一种同时检测多个靶序列的方法，该方法既快速简单，又灵敏。在这项工作中，我们描述了两个新的菁染料标记的dUTP类似物的合成，Cy3-dUTP和Cy5-dUTP。它们是DNA聚合酶的有效底物，可通过标准缺口平移、随机引发和聚合酶链式反应被结合到DNA探针中。确定了最佳的标记条件，每千碱基产生20-40个染料的探针。用这些类似物获得的直接标记的DNA探针为多色多序列分析提供了一种简单的方法，不需要二次检测试剂和步骤。
Fluorescence in situ hybridization (FISH) has become an indispensable tool in a variety of areas of research and clinical diagnostics. Many applications demand an approach for simultaneous detection of multiple target sequences that is rapid and simple, yet sensitive. In this work, we describe the synthesis of two new cyanine dye-labeled dUTP analogs, Cy3-dUTP and Cy5-dUTP. They are efficient substrates for DNA polymerases and can be incorporated into DNA probes by standard nick translation, random priming and polymerase chain reactions. Optimal labeling conditions have been identified which yield probes with 20-40 dyes per kilobase. The directly labeled DNA probes obtained with these analogs offer a simple approach for multicolor multisequence analysis that requires no secondary detection reagents and steps.