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Triphosphate reactive dyes can be used for enzymatic fluorescent labeling of DNA.
The fluorescent labeling method is a kind of non-radioactive method, which overcomes the safety, handling, stability, and cost problems caused by the radioactive labeling method, and is extensively used in the labeling and testing of nucleic acids, oligonucleotides, PCR products, and related molecules. checking. In the enzyme-mediated fluorescent labeling method, PCR uses DNA polymerases such as Taq and pfu to incorporate fluorescently-labeled primers or fluorescent molecule-modified nucleotides into the PCR products, which achieves the completion of the fluorescent labeling of the sample at the same time. Specific amplification of target fragments. dUTP is 2'-deoxyuridine 5'-triphosphate, which is often used in PCR, qPCR, RT-PCR, and other reactions to replace dTTP, so that the product obtained from the amplification reaction contains uracil.
Fluorescently labeled dUTP
Fluorescently labeled nucleotides are effective tools for detecting specific nucleic acid sequences in molecular biology and are usually combined with DNA or RNA sequences for detection and analysis through the action of enzymes.
Fluorescent molecule-modified dUTP could be used to produce fluorescently labeled DNA by nick translation, random primer extension, PCR, or end-tailing methods, without the use of dangerous and expensive radioisotope-labeled nucleotides. dUTP modified by fluorescent molecules is incorporated into DNA macromolecules through the action of enzymes. In this way, DNA is labeled with fluorescent molecules. The fluorescence quantum yield (Yf) produced by the combination of fluorescent molecules (such as cyanine) and nucleic acid molecules is significantly high and the detection sensitivity is also high. Fluorescently labeled dUTP has excellent safety, stability, and convenience.
dUTP can be modified by many various fluorescent molecules, so fluorescently labeled dUTP has a great range of potential applications. Fluorescently labeled dUTP is widely used to monitor PCR reactions, FISH probes, and microarray-based experiments.