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Synthesis and evaluation of a new non-fluorescent quencher in fluorogenic oligonucleotide probes for real-time PCR

Synthesis and evaluation of a new non-fluorescent quencher in fluorogenic oligonucleotide probes for real-time PCR
Synthesis and evaluation of a new non-fluorescent quencher in fluorogenic oligonucleotide probes for real-time PCR
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.
cystic-fibrosis gene, scorpion primers, deoxynucleoside phosphoramidites, molecular beacons, dna, identification, hybridization, fluorescence, polymerase, sequence
1477-0520
2534-2542
May, Jonathan P.
c820640d-d3d0-4eb9-b2f1-78d6ac689b8a
Brown, Lynda J.
75aa95fa-5d27-46a7-9dbe-0f465a664f5b
van Delft, Ian
0509bb2a-8b1a-43cc-b092-6c878d7ac91b
Thelwell, Nicola
f4f0808c-02a5-4380-a060-7537ad3b27ec
Harley, Kate
84ef368f-6263-45c9-909b-c8e06f6059fc
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
May, Jonathan P.
c820640d-d3d0-4eb9-b2f1-78d6ac689b8a
Brown, Lynda J.
75aa95fa-5d27-46a7-9dbe-0f465a664f5b
van Delft, Ian
0509bb2a-8b1a-43cc-b092-6c878d7ac91b
Thelwell, Nicola
f4f0808c-02a5-4380-a060-7537ad3b27ec
Harley, Kate
84ef368f-6263-45c9-909b-c8e06f6059fc
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89

May, Jonathan P., Brown, Lynda J., van Delft, Ian, Thelwell, Nicola, Harley, Kate and Brown, Tom (2005) Synthesis and evaluation of a new non-fluorescent quencher in fluorogenic oligonucleotide probes for real-time PCR. Organic & Biomolecular Chemistry, 3 (14), 2534-2542. (doi:10.1039/B504759E).

Record type: Article

Abstract

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.

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More information

e-pub ahead of print date: 7 July 2005
Published date: 2005
Keywords: cystic-fibrosis gene, scorpion primers, deoxynucleoside phosphoramidites, molecular beacons, dna, identification, hybridization, fluorescence, polymerase, sequence
Organisations: Chemistry

Identifiers

Local EPrints ID: 337199
URI: http://eprints.soton.ac.uk/id/eprint/337199
ISSN: 1477-0520
PURE UUID: e4b9030b-8032-44bc-b3e6-cfb0a6a3f358
ORCID for Lynda J. Brown: ORCID iD orcid.org/0000-0002-5678-0814

Catalogue record

Date deposited: 20 Apr 2012 12:46
Last modified: 29 Oct 2019 01:59

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Contributors

Author: Jonathan P. May
Author: Lynda J. Brown ORCID iD
Author: Ian van Delft
Author: Nicola Thelwell
Author: Kate Harley
Author: Tom Brown

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