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), 1-9. (doi:10.1093/nar/29.20.e96). (PMID:11600715).

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Description/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.

Item Type: Article
ISSNs: 0305-1048 (print)
1362-4962 (electronic)
Keywords: cystic-fibrosis gene, energy-transfer, dna, fluorescence, identification, hybridization, probes
Subjects: Q Science > QH Natural history > QH426 Genetics
R Medicine > RB Pathology
T Technology > TP Chemical technology
Divisions: University Structure - Pre August 2011 > School of Chemistry
ePrint ID: 19623
Date Deposited: 13 Feb 2006
Last Modified: 27 Mar 2014 18:09
Contact Email Address: tb2@soton.ac.uk
URI: http://eprints.soton.ac.uk/id/eprint/19623

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