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HyBeacon (TM) probes: a new tool for DNA sequence detection and allele discrimination

HyBeacon (TM) probes: a new tool for DNA sequence detection and allele discrimination
HyBeacon (TM) probes: a new tool for DNA sequence detection and allele discrimination
Technologies that permit rapid investigation of DNA sequences, such as those containing single nucleotide polymorphisms (SNPs), are of great consequence to many sectors that perform molecular diagnostic analyses. We have developed a novel fluorescent oligonucleoticle probe technology, termed HyBeacons(TM), which provides a new homogeneous method for fluorescence-based sequence detection, allele discrimination and DNA quantification. Hybridization of HyBeacons to complementary DNA target sequences results in a measurable elevation of probe fluorescence emission. HyBeacon probes may be incorporated into real-time polymerase chain reaction (PCR) assays to detect the presence and monitor the accumulation of specific DNA sequences. Furthermore, closely related sequences differing by as little as a single nucleotide maybe discriminated by measuring the melting temperatures (T-m) of various probe/target duplexes and exploiting the differences in T-m that exist between different duplexes. We demonstrate here that HyBeacon probes are efficient tools for rapid sequence analysis and that a single probe may be employed to reliably identify homozygous and heterozygous samples. Additional benefits exhibited by the HyBeacon technology derive from their simple mode of action, ease of design, relatively inexpensive synthesis and potential for multiplex analysis.
homogeneous analysis, luorescent probes, lightcycler, single nucleotide polymorphisms, allele discriminationcystic-fibrosis gene, light-up probes, factor-v-leiden, molecularbeacons, energy-transfer, fluorescence properties, -nuclease assay, point mutation, melting curves, nucleic-acid
0890-8508
363-374
French, D.J.
e7221974-9b57-44a5-9cfb-1260526131fa
Archard, C.L.
4069d8a6-f914-4d2c-97cc-af0a3427ee7d
Brown, T.
a64aae36-bb30-42df-88a2-11be394e8c89
McDowell, D.G.
30e07793-9137-40ea-90ca-7752ddd2e7e7
French, D.J.
e7221974-9b57-44a5-9cfb-1260526131fa
Archard, C.L.
4069d8a6-f914-4d2c-97cc-af0a3427ee7d
Brown, T.
a64aae36-bb30-42df-88a2-11be394e8c89
McDowell, D.G.
30e07793-9137-40ea-90ca-7752ddd2e7e7

French, D.J., Archard, C.L., Brown, T. and McDowell, D.G. (2001) HyBeacon (TM) probes: a new tool for DNA sequence detection and allele discrimination. Molecular and Cellular Probes, 15 (6), 363-374. (doi:10.1006/mcpr.2001.0384).

Record type: Article

Abstract

Technologies that permit rapid investigation of DNA sequences, such as those containing single nucleotide polymorphisms (SNPs), are of great consequence to many sectors that perform molecular diagnostic analyses. We have developed a novel fluorescent oligonucleoticle probe technology, termed HyBeacons(TM), which provides a new homogeneous method for fluorescence-based sequence detection, allele discrimination and DNA quantification. Hybridization of HyBeacons to complementary DNA target sequences results in a measurable elevation of probe fluorescence emission. HyBeacon probes may be incorporated into real-time polymerase chain reaction (PCR) assays to detect the presence and monitor the accumulation of specific DNA sequences. Furthermore, closely related sequences differing by as little as a single nucleotide maybe discriminated by measuring the melting temperatures (T-m) of various probe/target duplexes and exploiting the differences in T-m that exist between different duplexes. We demonstrate here that HyBeacon probes are efficient tools for rapid sequence analysis and that a single probe may be employed to reliably identify homozygous and heterozygous samples. Additional benefits exhibited by the HyBeacon technology derive from their simple mode of action, ease of design, relatively inexpensive synthesis and potential for multiplex analysis.

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More information

Published date: 1 December 2001
Keywords: homogeneous analysis, luorescent probes, lightcycler, single nucleotide polymorphisms, allele discriminationcystic-fibrosis gene, light-up probes, factor-v-leiden, molecularbeacons, energy-transfer, fluorescence properties, -nuclease assay, point mutation, melting curves, nucleic-acid

Identifiers

Local EPrints ID: 19480
URI: http://eprints.soton.ac.uk/id/eprint/19480
ISSN: 0890-8508
PURE UUID: 4c0c9dbf-ede4-423e-95a2-ec5a91c6afc0

Catalogue record

Date deposited: 16 Feb 2006
Last modified: 15 Jul 2019 19:27

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