DNA SEQUENCING BY SYNTHESIS USING RAMAN AND INFRARED SPECTROSCOPY DETECTION
First Claim
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1. A nucleoside triphosphate analogue having the structure:
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Abstract
This invention provides nucleoside triphosphate analogues having the structure:
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- wherein B is a base and is adenine, guanine, cytosine, uracil or thymine, wherein R″ is an OH or an H, and wherein R′ is azidomethyl, a hydrocarbyl, or a substituted hydrocarbyl, and which has a Raman spectroscopy peak with wavenumber from 2000 cm−1 to 2300 cm−1 or a Fourier transform-infrared spectroscopy spectroscopy peak with wavenumber from 2000 cm−1 to 2300 cm−1, and also to methods of DNA sequencing and SNP detection.
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Citations
78 Claims
- 1. A nucleoside triphosphate analogue having the structure:
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2-5. -5. (canceled)
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7. A polynucleotide analogue, wherein the polynucleotide analogue differs from a polynucleotide by comprising at its 3′
- terminus one of the following structures in place of the H atom of the 3′
OH group normally present in the polynucleotide;
- View Dependent Claims (26, 29, 30)
- terminus one of the following structures in place of the H atom of the 3′
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11. (canceled)
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14. (canceled)
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16-19. -19. (canceled)
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21. A nucleoside triphosphate analogue having the structure:
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22-25. -25. (canceled)
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27-28. -28. (canceled)
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31-40. -40. (canceled)
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43. A method for determining the identity of a nucleotide residue within a stretch of consecutive nucleic acid residues in a single-stranded DNA comprising:
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(a) contacting the single-stranded DNA with four different oligonucleotide probes, (1) wherein each of the oligonucleotide probes comprises (i) a portion that is complementary to a portion of consecutive nucleotides of the single stranded DNA immediately 3′
to the nucleotide residue being identified, and (ii) a 3′
terminal nucleotide residue analogue comprising on its sugar a 3′
-O—
R′
group wherein R′
is (a) is azidomethyl, or a substituted or unsubstituted hydrocarbyl group and (b) has a predetermined Raman spectroscopy peak with wavenumber which is from 2000 cm−
1 to 2300 cm−
1, and which is different from the wavenumber of the Raman spectroscopy peak of the R′
of the 3′
terminal nucleotide residue analogue of the other three oligonucleotide probes, or has a predetermined Fourier-transform infra red spectroscopy peak with wavenumber which is from 2000 cm−
1 to 2300 cm−
1, and is different from the wavenumber of the Fourier-transform infra red spectra peak of the R′
of the 3′
terminal nucleotide residue analogue of the other three oligonucleotide probes, and (iii) each of the four terminal nucleotide residue analogue comprises a base which is different from the base of the terminal nucleotide residue analogue of the other three oligonucleotide probes, and (2) under conditions permitting hybridization of the primer which is fully complementary to the portion of consecutive nucleotides of the single stranded DNA immediately 3′
to the nucleotide residue being identified;(b) removing oligonucleotide primers not hybridized to the single-stranded DNA; and (c) determining the wavenumber of the Raman spectroscopy peak or wavenumber of the Fourier-transform infra red peak of the dNTP analogue of the oligonucleotide probe hybridized in step (a) so as to thereby determine the identity of the dNTP analogue of the hybridized oligonucleotide probe and thus determine the identity of the complementary nucleotide residue in the single-stranded DNA, thereby identifying the nucleotide residue within the stretch of consecutive nucleic acid residues in the single-stranded DNA. - View Dependent Claims (78)
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44-46. -46. (canceled)
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48-76. -76. (canceled)
Specification