Synthesis and evaluation of a new non-fluorescent quencher in fluorogenic oligonucleotide probes for real-time PCR
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).
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Description/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.
| Item Type: | Article |
|---|---|
| ISSNs: | 1477-0520 (print) 1477-0539 (electronic) |
| Keywords: | cystic-fibrosis gene, scorpion primers, deoxynucleoside phosphoramidites, molecular beacons, dna, identification, hybridization, fluorescence, polymerase, sequence |
| Subjects: | Q Science > QD Chemistry Q Science > QH Natural history > QH426 Genetics |
| Divisions: | Faculty of Natural and Environmental Sciences > Chemistry |
| Item ID: | 337199 |
| Date Deposited: | 20 Apr 2012 12:46 |
| Last Modified: | 26 Apr 2013 05:42 |
| Contributors: | May, Jonathan P. (Author) Brown, Lynda J. (Author) van Delft, Ian (Author) Thelwell, Nicola (Author) Harley, Kate (Author) Brown, Tom (Author) |
| Date: | 2005 |
| Status: | Published |
| URI: | http://eprints.soton.ac.uk/id/eprint/337199 |
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