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A non-fluorescent quencher, based on the diaminoanthraquinone Disperse Blue 3, has been incorporated into oligonucleotides at the 5'-end, the 3'-end and internally as a thymidine derivative. Fluorimetry and fluorogenic real-time PCR experiments demonstrate that the quencher is effective with a wide range of fluorescent dyes. The anthraquinone moiety increases the melting temperature of DNA duplexes, thus allowing shorter, more discriminatory probes to be used. The quencher has been used in Scorpion primers and TaqMan probes for human DNA sequence recognition and mutation detection.

Original publication

DOI

10.1039/b504759e

Type

Journal article

Journal

Org Biomol Chem

Publication Date

21/07/2005

Volume

3

Pages

2534 - 2542

Keywords

Azo Compounds, Base Sequence, Chromogenic Compounds, Fluorescent Dyes, Molecular Sequence Data, Oligonucleotide Probes, Organophosphorus Compounds, Polymerase Chain Reaction, Spectrum Analysis