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Duplex Scorpion primers in SNP analysis and FRET applications

Duplex Scorpion primers in SNP analysis and FRET applications
Duplex Scorpion primers in SNP analysis and FRET applications
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.
cystic-fibrosis gene, energy-transfer, dna, fluorescence, identification, hybridization, probes
0305-1048
1-9
Solinas, Antonio
a3843e95-3cf0-499e-b264-38a2a8ba91eb
Brown, Lynda J.
75aa95fa-5d27-46a7-9dbe-0f465a664f5b
McKeen, Catherine
7bcc5905-63fc-4007-a88c-ea967e7ee7e5
Mellor, John M.
116e0a09-9f67-4a1e-8110-9a6f27772ed3
Nicol, Jamie T.G.
eb81f29d-a3e0-47d9-bec2-1f254ee372c4
Thelwell, Nicky
c94f6f0c-3fd0-4d68-b32a-3e4e2f47afab
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Solinas, Antonio
a3843e95-3cf0-499e-b264-38a2a8ba91eb
Brown, Lynda J.
75aa95fa-5d27-46a7-9dbe-0f465a664f5b
McKeen, Catherine
7bcc5905-63fc-4007-a88c-ea967e7ee7e5
Mellor, John M.
116e0a09-9f67-4a1e-8110-9a6f27772ed3
Nicol, Jamie T.G.
eb81f29d-a3e0-47d9-bec2-1f254ee372c4
Thelwell, Nicky
c94f6f0c-3fd0-4d68-b32a-3e4e2f47afab
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89

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)

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.

Full text not available from this repository.

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
ORCID for Lynda J. Brown: ORCID iD orcid.org/0000-0002-5678-0814

Catalogue record

Date deposited: 13 Feb 2006
Last modified: 17 Dec 2019 01:55

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