CYANINE DYE DUTP ANALOGS FOR ENZYMATIC LABELING OF DNA PROBES

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 wor k, we describe the synthesis of two new cyanine dye-labeled dUTP analo gs, Cy3-dUTP and Cy5-dUTP. They are efficient substrates for DNA polym erases and can be incorporated into DNA probes by standard nick transl ation, random priming and polymerase chain reactions. Optimal labeling conditions have been identified which yield probes with 20-40 dyes pe r kilobase. The directly labeled DNA probes obtained with these analog s offer a simple approach for multicolor multisequence analysis that r equires no secondary detection reagents and steps.