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Duplex Scorpion primers in SNP analysis and FRET applications

Solinas, Antonio, Brown, Lynda J., McKeen, Catherine, Mellor, John M., Nicol, Jamie T.G., Thelwell, Nicky and Brown, Tom (2001) Duplex Scorpion primers in SNP analysis and FRET applications Nucleic Acids Research, 29, (20), pp. 1-9. (doi:10.1093/nar/29.20.e96). (PMID:11600715).

Record type: Article

Abstract

Scorpions are fluorogenic PCR primers with a probe element attached at the 5'-end via a PCR stopper. They are used in real-time amplicon-specific detection of PCR products in homogeneous solution. Two different formats are possible, the ‘stem–loop’ format and the ‘duplex’ format. In both cases the probing mechanism is intramolecular.

We have shown that duplex Scorpions are efficient probes in real-time PCR. They give a greater fluorescent signal than stem–loop Scorpions due to the vastly increased separation between fluorophore and quencher in the active form. We have demonstrated their use in allelic discrimination at the W1282X locus of the ABCC7 gene and shown that they can be used in assays where fluorescence resonance energy transfer is required.

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More information

Published date: 15 October 2001
Keywords: cystic-fibrosis gene, energy-transfer, dna, fluorescence, identification, hybridization, probes

Identifiers

Local EPrints ID: 19623
URI: http://eprints.soton.ac.uk/id/eprint/19623
ISSN: 0305-1048
PURE UUID: 6a20bea9-98ed-4cbb-ad4a-be575ce9ecdf

Catalogue record

Date deposited: 13 Feb 2006
Last modified: 17 Jul 2017 16:30

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Contributors

Author: Antonio Solinas
Author: Lynda J. Brown
Author: Catherine McKeen
Author: John M. Mellor
Author: Jamie T.G. Nicol
Author: Nicky Thelwell
Author: Tom Brown

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