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High throughput measurement of duplex, triplex and quadruplex melting curves using molecular beacons and a LightCycler

Darby, R.A.J., Sollogoub, M., McKeen, M., Brown, Lynda J., Risitano, A., Brown, N., Barton, C., Brown, Tom and Fox, K.R. (2002) High throughput measurement of duplex, triplex and quadruplex melting curves using molecular beacons and a LightCycler Nucleic Acids Research, 30, (9), e39-[9pp]. (doi:10.1093/nar/30.9.e39). (PMID:11972354).

Record type: Article


We have used oligonucleotides containing molecular beacons to determine melting profiles for intramolecular DNA duplexes, triplexes and quadruplexes (tetraplexes). The synthetic oligonucleotides used in these studies contain a fluorophore (fluorescein) and quencher (methyl red) attached either to deoxyribose or to the 5 position of dU. In the folded DNA structures the fluorophore and quencher are in close proximity and the fluorescence is quenched. When the structures melt, the fluorophore and quencher are separated and there is a large increase in fluorescence. These experiments were performed in a Roche LightCycler; this requires small amounts of material (typically 4 pmol oligonucleotide) and can perform 32 melting profiles in parallel. We have used this technique to compare the stability of triplexes containing different base analogues and to confirm the selectivity of a triplex-binding ligand for triplex, rather than duplex, DNA. We have also compared the melting of inter- and intramolecular quadruplexes.

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Submitted date: 8 February 2002
Published date: 1 May 2002
Keywords: resonance energy-transfer, double-stranded dna, g-quartet formation, 2', -aminoethoxy-modified oligonucleotides, dual recognition, scorpionprimers, promoter region, helix formation, nucleic-acids, fluorescence


Local EPrints ID: 24241
ISSN: 0305-1048
PURE UUID: e90f1ce0-8489-43eb-a725-3577954baa3a
ORCID for K.R. Fox: ORCID iD

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Date deposited: 29 Mar 2006
Last modified: 17 Jul 2017 16:14

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Author: R.A.J. Darby
Author: M. Sollogoub
Author: M. McKeen
Author: Lynda J. Brown
Author: A. Risitano
Author: N. Brown
Author: C. Barton
Author: Tom Brown
Author: K.R. Fox ORCID iD

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