Cyanine dye dUTP analogs for enzymatic labeling of DNA probes.
AUTOR(ES)
Yu, H
RESUMO
Fluorescence in situ hybridization (FISH) has become and 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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=310300Documentos Relacionados
- Ferrocene conjugates of dUTP for enzymatic redox labelling of DNA
- Detection of DNA Damage in Prokaryotes by Terminal Deoxyribonucleotide Transferase-Mediated dUTP Nick End Labeling
- Archaeal dUTPase enhances PCR amplifications with archaeal DNA polymerases by preventing dUTP incorporation
- Human dUTP pyrophosphatase: cDNA sequence and potential biological importance of the enzyme.
- Rapid detection of DNA-binding factors using protein-blotting and digoxigenin-dUTP marked probes.