Back to results
US 6,475,730 B1Granted
Detecting nucleic acids
AssigneeThe Victoria University of Manchester
InventorsKenneth T Douglas, Elena V Bichenkova
Life of this patent
Estimating the term…
Abstract
The present invention relates to a method of detecting nucleic acids and particularly to a method of analyzing for the presence and/or amount of a nucleic acid.
Claims47
A method of analysing a sample to determine the presence or otherwise therein of a target polynucleotide sequence, the method comprising
- (a) treating the sample under hybridizing conditions with
- (i) a first polynucleotide probe having a 5′-terminal nucleotide labelled with a first chromophoric moiety able on photo-irradiation to form a complex with a second chromophoric moiety, and
- (ii) a second polynucleotide probe labelled at its 3′-terminal nucleotide with moiety, said first and second probes being adapted to bind to mutually exclusive regions of said target sequence such that said moieties are at the proximal ends of the probes and are able to form said complex which is detectably different from the first and second moieties,
- (b) effecting photo-irradiation to cause complex formation, and
- (c) detecting for formation of the complex, characterised in that
- (d) said first and second moieties are different from each other and are capable of forming an exciplex as the detectable complex, and
- (e) the first and second moieties come into exciplex forming relationship in a localised region of greater hydrophobicity than the bulk phase of the sample being analysed.
4 more elements
A method of analysing a sample to determine the presence or otherwise therein of a target polynucleotide sequence, the method comprising
- (a) treating the sample under hybridizing conditions with
- (i) a first polynucleotide probe having a 5′-terminal nucleotide labelled with a first chromophoric moiety able on photo-irradiation to form a complex with a second chromophoric moiety, and
- (ii) a second polynucleotide probe labelled at its 3′-terminal nucleotide with moiety, said first and second probes being adapted to bind to mutually exclusive regions of said target sequence such that said moieties are at the proximal ends of the probes and are able to form said complex which is detectably different from the first and second moieties,
- (b) effecting photo-irradiation to cause complex formation, and
- (c) detecting for formation of the complex, characterised in that
- (d) said first and second moieties are different from each other and are capable of forming an exciplex as the detectable complex, and
- (e) the first and second moieties come into exciplex forming relationship within the immediate vicinity of the double stranded nucleic acid.
4 more elements
A method of analysing a sample to determine the presence or otherwise therein of a target polynucleotide sequence, the method comprising
- (a) treating the sample under hybridizing conditions with
- (i) a first polynucleotide probe having a 5′-terminal nucleotide labelled with a first chromophoric moiety able on photo-irradiation to form a complex with a second chromophoric moiety, and
- (ii) a second polynucleotide probe labelled at its 3′-terminal nucleotide with moiety, said first and second probes being adapted to bind to mutually exclusive regions of said target sequence such that said moieties are at the proximal ends of the probes and are able to form said complex which is detectably different from the first and second moieties,
- (b) effecting photo-irradiation to cause complex formation, and
- (c) detecting for formation of the complex, characterised in that
- (d) said first and second moieties are different from each other and are capable of forming an exciplex as the detectable complex, and
- (e) there is included in the sample being analysed a compound capable of developing an exciplex fluorescent signal when the first and second moieties are in exciplex forming relationship.
4 more elements
A method of analysing a sample to determine the presence or otherwise therein of a target polynucleotide sequence, the method comprising
- (a) treating the sample under hybridizing conditions with
- (i) a first polynucleotide probe having a 5′-terminal nucleotide labelled with a first chromophoric moiety able on photo-irradiation to form a complex with a second chromophoric moiety, and
- (ii) a second polynucleotide probe labelled at its 3′-terminal nucleotide with moiety, said first and second probes being adapted to bind to mutually exclusive regions of said target sequence such that said moieties are at the proximal ends of the probes and are able to form said complex which is detectably different from the first and second moieties,
- (b) effecting photo-irradiation to cause complex formation, and
- (c) detecting for formation of the complex, characterised in that
- (d) said first and second moieties are different from each other and are capable of forming an exciplex as the detectable complex, and
- (e) a magnetic field is applied to enhance exciplex emission.
4 more elements
A method of analysing a sample to determine the presence or otherwise therein of a target polynucleotide sequence, the method comprising
- (a) treating the sample under hybridizing conditions with
- (i) a first polynucleotide probe having a 5′-terminal nucleotide labelled with a first chromophoric moiety able on photo-irradiation to form a complex with a second chromophoric moiety, and
- (ii) a second polynucleotide probe labelled at its 3′-terminal nucleotide with moiety, said first and second probes being adapted to bind to mutually exclusive regions of said target sequence such that said moieties are at the proximal ends of the probes and are able to form said complex which is detectably different from the first and second moieties,
- (b) effecting photo-irradiation to cause complex formation, and
- (c) detecting for formation of the complex, characterised in that
- (d) said first and second moieties are different from each other and are capable of forming an exciplex as the detectable complex, and in that
- (e) the method involves at least on of the steps, in either order, of (i) at least partial removal of the relatively polar medium, and (ii) addition of a less polar medium
- thereby to enhance exciplex emission.
5 more elements
Dependencies are inferred from the claim text.