Nucleic acid sequencing by raman monitoring of molecular deconstruction
First Claim
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1. ) An apparatus comprising:
- a) a reaction chamber to contain an immobilization surface, the immobilization surface to attach a single nucleic acid molecule;
b) a flow path attached to the reaction chamber; and
c) a detection unit comprising an excitation source and a Raman spectroscopic detector.
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Abstract
The disclosed methods, apparatus and compositions are of use for nucleic acid sequencing. More particularly, the methods and apparatus concern sequencing single molecules of single stranded DNA or RNA by exposing the molecule to exonuclease activity, removing free nucleotides one at a time from one end of the nucleic acid, and identifying the released nucleotides by Raman spectroscopy or FRET.
76 Citations
28 Claims
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1. ) An apparatus comprising:
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a) a reaction chamber to contain an immobilization surface, the immobilization surface to attach a single nucleic acid molecule;
b) a flow path attached to the reaction chamber; and
c) a detection unit comprising an excitation source and a Raman spectroscopic detector. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. ) An apparatus comprising:
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a) a reaction chamber to contain an immobilization surface, the immobilization surface to attach a single nucleic acid molecule; and
b) a detection unit comprising an excitation source and a Raman spectroscopic detector. - View Dependent Claims (11, 12, 13, 14, 15, 16)
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17. ) A method of sequencing nucleic acids comprising:
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a) preparing a single nucleic acid molecule, one end of the nucleic acid molecule attached to an immobilization surface within a reaction chamber;
b) sequentially removing nucleotides from the unattached end of the nucleic acid molecule;
c) separating the nucleotides from the nucleic acid molecule; and
d) detecting the nucleotides by Raman spectroscopy. - View Dependent Claims (18, 19, 20)
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21. ) A method of sequencing nucleic acids comprising:
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a) preparing a single nucleic acid molecule, one end of the nucleic acid molecule attached to an immobilization surface within a reaction chamber;
b) sequentially removing nucleotides from the unattached end of the nucleic acid molecule; and
c) quantifying the nucleotides in the reaction chamber by Raman spectroscopy. - View Dependent Claims (22, 23, 24)
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25. ) A method of sequencing nucleic acids comprising:
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a) preparing a single nucleic acid molecule, one end of the nucleic acid molecule attached to an immobilization surface within a reaction chamber;
b) sequentially removing nucleotides from the unattached end of the nucleic acid molecule; and
c) detecting the nucleotides by fluorescence resonance energy transfer (FRET) spectroscopy. - View Dependent Claims (26, 27, 28)
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Specification