Nucleotide analogs for sequencing
First Claim
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1. A nucleotide analog comprising:
- a) a nucleotide comprising a phosphate moiety, a base, and a sugar;
b) a photocleavable terminating moiety attached to the nucleotide by a first photocleavable linker, wherein said photocleavable terminating moiety is electrochemically detectable by accepting or donating at least one electron during an electrochemical reaction; and
c) a second photocleavable terminating moiety attached to the nucleotide by a second photocleavable linker.
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Abstract
Provided herein is technology relating to sequencing nucleic acids, but not exclusively, to compositions, methods, systems, and kits related to nucleotides comprising an electrochemically detectable moiety and one or more photolabile synthesis-inhibiting moieties.
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12 Claims
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1. A nucleotide analog comprising:
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a) a nucleotide comprising a phosphate moiety, a base, and a sugar; b) a photocleavable terminating moiety attached to the nucleotide by a first photocleavable linker, wherein said photocleavable terminating moiety is electrochemically detectable by accepting or donating at least one electron during an electrochemical reaction; and c) a second photocleavable terminating moiety attached to the nucleotide by a second photocleavable linker. - View Dependent Claims (2, 3, 4, 10, 11, 12)
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5. A method for sequencing a nucleic acid, the method comprising:
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1) hybridizing a primer to a nucleic acid to form a hybridized primer/nucleic acid complex; 2) providing a plurality of nucleotide analogs, each nucleotide analog comprising a nucleotide and a photocleavable terminating moiety attached to the nucleotide, wherein said photocleavable terminating moiety is electrochemically detectable by accepting or donating at least one electron during an electrochemical reaction; 3) reacting the hybridized primer/nucleic acid complex and the nucleotide analog with a polymerase to add the nucleotide analog to the primer by a polymerase reaction to form an extended product comprising an incorporated nucleotide analog; 4) querying the extended product to identify the incorporated nucleotide analog; 5) exposing the extended product to a light source providing a first wavelength of light to remove an electrochemically detectable, photocleavable terminating moiety from the incorporated nucleotide analog; and wherein each nucleotide analog further comprises a second photocleavable terminating moiety attached to the nucleotide and the method further comprises exposing the extended product to a light source providing a second wavelength of light to remove a second photocleavable terminating moiety from the incorporated nucleotide analog. - View Dependent Claims (6, 7, 8, 9)
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Specification