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Mode of action and application of Scorpion primers to mutation detection

Record type: Article

Scorpion primers can be used to detect PCR products in homogeneous solution, Their structure promotes a unimolecular probing mechanism. We compare their performance with that of the same probe sequence forced to act in a bimolecular manner. The data suggest that Scorpions indeed probe by a unimolecular mechanism which is faster and move efficient than the bimolecular mechanism. This mechanism is not dependent on enzymatic cleavage of the probe. A direct comparison between Scorpions, TaqMan and Molecular Beacons on a Roche Light-Cycler indicates that Scorpions perform better, particularly under fast cycling conditions, Development of a cystic fibrosis mutation detection assay shows that Scorpion primers are selective enough to detect single base mutations and give good sensitivity in all cases, Simultaneous detection of both normal and mutant alleles in a single reaction is possible by combining two Scorpions in a multiplex reaction. Such favourable properties of Scorpion primers should make the technology ideal in numerous applications.

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Citation

Thelwell, N., Millington, S., Solinas, A., Booth, J. and Brown, T. (2000) Mode of action and application of Scorpion primers to mutation detection Nucleic Acids Research, 28, (19), pp. 3752-3761. (doi:10.1093/nar/28.19.3752).

More information

Published date: 1 October 2000
Keywords: cystic-fibrosis gene, molecular beacons, cftr gene, identification, dna, fluorescence

Identifiers

Local EPrints ID: 18970
URI: http://eprints.soton.ac.uk/id/eprint/18970
ISSN: 0305-1048
PURE UUID: 800c65a5-8f84-45a0-a1c3-dba806ef0bb0

Catalogue record

Date deposited: 19 Jan 2006
Last modified: 17 Jul 2017 16:34

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Contributors

Author: N. Thelwell
Author: S. Millington
Author: A. Solinas
Author: J. Booth
Author: T. Brown

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