Detection of nucleic acid polymorphisms
First Claim
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1. A method for characterizing nucleic acid polymorphisms, including the steps:
- (a) provision of a nucleic acid template to be investigated, (b) annealing of at least one starting primer onto the nucleic acid template, with the 3′
end of the starting primer being located upstream of a nucleic acid polymorphism to be investigated, (c) extension of the starting primer with at least one fluorescence-labeled nucleotide and (d) detection of nucleotides incorporated into the starting primer by single-molecule determination.
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Abstract
Methods are described methods functioning at the single-molecule level for characterizing nucleotide polymorphisms.
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33 Claims
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1. A method for characterizing nucleic acid polymorphisms, including the steps:
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(a) provision of a nucleic acid template to be investigated, (b) annealing of at least one starting primer onto the nucleic acid template, with the 3′
end of the starting primer being located upstream of a nucleic acid polymorphism to be investigated,(c) extension of the starting primer with at least one fluorescence-labeled nucleotide and (d) detection of nucleotides incorporated into the starting primer by single-molecule determination. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 25, 26, 27, 28, 29, 30, 31)
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20. A method for characterizing nucleic acid polymorphisms in a microwell, including the steps:
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(a) provision of a carrier particle with a nucleic acid molecule which is immobilized thereon and consists of a single-stranded nucleic acid template and of a starting primer, (b) extension of the starting primer by a fluorescence-labeled chain-termination molecule, (c) where appropriate washing of the well to remove unincorporated labels and (d) detection of the fluorescent label incorporated into the starting primer. - View Dependent Claims (21, 22, 23, 24)
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- 32. A method for increasing the detection efficiency in the detection of the fluorescence from single molecules, characterized in that a dispersion element is used to separate the light of different wavelengths.
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