DNA Sequencing by Nanopore Using Modified Nucleotides
First Claim
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1. A method for determining the nucleotide sequence of a single-stranded DNA comprising the steps of:
- (a) passing the single-stranded DNA through a pore of suitable diameter by applying an electric field to the DNA, wherein at least each A or each G residue and at least each C, each T or each U residue comprises a modifying group bound to its respective base so that each type of nucleotide in the DNA has an electronic signature which is distinguishable from the electronic signature of each other type of nucleotide in the DNA;
(b) for each nucleotide of the DNA which passes through the pore, determining an electronic signature for such nucleotide; and
(c) comparing each electronic signature determined in step (b) with electronic signatures corresponding to each of A, G, C and T modified as per the nucleotides in the single-stranded DNA, so as to determine the identity of each such nucleotide, thereby determining the nucleotide sequence of the single-stranded DNA.
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Abstract
This invention provides a process for sequencing single-stranded DNA by employing a nanopore and modified nucleotides.
195 Citations
21 Claims
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1. A method for determining the nucleotide sequence of a single-stranded DNA comprising the steps of:
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(a) passing the single-stranded DNA through a pore of suitable diameter by applying an electric field to the DNA, wherein at least each A or each G residue and at least each C, each T or each U residue comprises a modifying group bound to its respective base so that each type of nucleotide in the DNA has an electronic signature which is distinguishable from the electronic signature of each other type of nucleotide in the DNA; (b) for each nucleotide of the DNA which passes through the pore, determining an electronic signature for such nucleotide; and (c) comparing each electronic signature determined in step (b) with electronic signatures corresponding to each of A, G, C and T modified as per the nucleotides in the single-stranded DNA, so as to determine the identity of each such nucleotide, thereby determining the nucleotide sequence of the single-stranded DNA. - View Dependent Claims (3, 4, 5, 6, 8, 9, 11)
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2. A method for determining the nucleotide sequence of a single-stranded RNA comprising the steps of:
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(a) passing the single-stranded RNA through a pore of suitable diameter by applying an electric field to the RNA, wherein at least each A or each G residue and at least each C or each U residue comprises a modifying group bound to its respective base so that each type of nucleotide in the RNA has an electronic signature which is distinguishable from the electronic signature of each other type of nucleotide in the RNA; (b) for each nucleotide of the RNA which passes through the pore, determining an electronic signature for such nucleotide; and (c) comparing each electronic signature determined in step (b) with electronic signatures corresponding to each of A, G, C and U modified as per the nucleotides in the single-stranded RNA, so as to determine the identity of each such nucleotide, thereby determining the nucleotide sequence of the single-stranded RNA. - View Dependent Claims (7, 10, 12, 18, 19, 20, 21)
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- 13. A nucleotide having an azido group covalently bound to its base.
Specification