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PMID: 11600715 Published · ppublish English Comparative Study Evaluation Study Journal Article Research Support, Non-U.S. Gov't

Duplex Scorpion primers in SNP analysis and FRET applications.

Nucleic acids research ·Vol. 29 ·No. 20 ·2001-10-15 ·Pages E96

Solinas A, Brown LJ, McKeen C, Mellor JM, Nicol J, Thelwell N, Brown T

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.

MeSH Terms

DNA Primers Energy Transfer Fluorescence Polymerase Chain Reaction/methods Temperature

Chemicals

DNA Primers

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Solinas A

Department of Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK.

Brown L J

McKeen C

Mellor J M

Nicol J

Thelwell N

Brown T

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14 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

1362-4962

DOI

10.1093/nar/29.20.e96

Published

2001-10-15

Pages

E96

Language

English

Region

England

NLM ID

0411011

PMCID

PMC60224

Subset

External Links

PubMed source DOI: 10.1093/nar/29.20.e96 PMC Full Text View journal details

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