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Synthesis and properties of new fluorescent dyes and dye-labelled oligonucleotides

Synthesis and properties of new fluorescent dyes and dye-labelled oligonucleotides
Synthesis and properties of new fluorescent dyes and dye-labelled oligonucleotides
Fluorescence is widely used in DNA sequencing and genetic analysis. Novel fluorescence techniques for studying nucleic acids and other biologically important molecules are constantly being developed. In order to provide efficient probes with desired fluorescent properties, new fluorophores and nucleic acids labelling chemistries are required. The synthesis of various cyanine dyes derivatives is described in this thesis. Dyes were designed for NHS-ester and click chemistry labelling of oligonucleotides. Additionally the synthesis of disperse blue quencher and a 2?-mesyl-modified thymidine monomer are also presented.

A simple and convenient method of oligonucleotide labelling was developed. This method utilises the copper-catalysed 1,3-dipolar cycloaddition reaction (known as click chemistry) between alkyne-modified cyanine dyes and azide modified oligonucleotides. The conditions of the labelling were optimised and have proved efficient for solid-phase and solution-phase labelling of oligonucleotides with alkyne-functionalised dyes.

Click labelling was applied for the preparation of a series of HyBeacon probes and their fluorescence properties were investigated. The click-labelled HyBeacons were compared with HyBeacons labelled with standard cyanine dye NHS-esters showing improvements in the properties of the probes labelled by click chemistry.

The thesis also contains a detailed discussion of the photophysical properties of free cyanine dyes in solution and while attached to oligonucleotide probes.
Gerowska, Marta
e8cf7fc7-8111-4eff-aa0c-c576b59a9682
Gerowska, Marta
e8cf7fc7-8111-4eff-aa0c-c576b59a9682
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89

(2012) Synthesis and properties of new fluorescent dyes and dye-labelled oligonucleotides. University of Southampton, Chemistry, Doctoral Thesis, 308pp.

Record type: Thesis (Doctoral)

Abstract

Fluorescence is widely used in DNA sequencing and genetic analysis. Novel fluorescence techniques for studying nucleic acids and other biologically important molecules are constantly being developed. In order to provide efficient probes with desired fluorescent properties, new fluorophores and nucleic acids labelling chemistries are required. The synthesis of various cyanine dyes derivatives is described in this thesis. Dyes were designed for NHS-ester and click chemistry labelling of oligonucleotides. Additionally the synthesis of disperse blue quencher and a 2?-mesyl-modified thymidine monomer are also presented.

A simple and convenient method of oligonucleotide labelling was developed. This method utilises the copper-catalysed 1,3-dipolar cycloaddition reaction (known as click chemistry) between alkyne-modified cyanine dyes and azide modified oligonucleotides. The conditions of the labelling were optimised and have proved efficient for solid-phase and solution-phase labelling of oligonucleotides with alkyne-functionalised dyes.

Click labelling was applied for the preparation of a series of HyBeacon probes and their fluorescence properties were investigated. The click-labelled HyBeacons were compared with HyBeacons labelled with standard cyanine dye NHS-esters showing improvements in the properties of the probes labelled by click chemistry.

The thesis also contains a detailed discussion of the photophysical properties of free cyanine dyes in solution and while attached to oligonucleotide probes.

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Published date: 30 September 2012
Organisations: University of Southampton, Chemistry

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Local EPrints ID: 348171
URI: http://eprints.soton.ac.uk/id/eprint/348171
PURE UUID: 30c8cb85-a3a1-49b8-b161-2c5ccf9b9030

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Date deposited: 28 Feb 2013 14:54
Last modified: 18 Jul 2017 04:53

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