Sequence determination by direct detection
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Abstract
A sequencing methodology is disclosed that allows a single DNA or RNA molecule or portion thereof to be sequenced directly and in substantially real time. The methodology involves engineering a polymerase and/or dNTPs with atomic and/or molecular tags that have a detectable property that is monitored by a detection system.
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Citations
49 Claims
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31. A method for sequencing a molecular sequence comprising:
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supplying a sequence of nucleotides or nucleotide analogs to a sequencer comprising a polymerase and monomer types for the polymerase, each monomer type having a unique fluorescent tag covalently bonded a portion of the monomer that is released after incorporation of the monomer by the polymerase;
exciting the fluorescent tag with light from an excitation light source;
detecting a property of emitted fluorescent light from a tag before, during and/or after monomer incorporation via a fluorescent light detector; and
converting the detected property into an identity of the incorporated monomer complementary to a nucleotide in the sequence. - View Dependent Claims (35, 36, 37, 38, 39, 40, 41)
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42. A method for sequencing a molecular sequence comprising:
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supplying a sequence of nucleotides or nucleotide analogs to a sequencer comprising a polymerase and monomer types for the polymerase, each monomer type having a unique fluorescent tag covalently bonded to a portion of the monomer that is released after incorporation of the monomer by the polymerase;
exciting the fluorescent tag with light from an excitation light source;
detecting a property of emitted fluorescent light before, during and/or after a series of monomer incorporations via a fluorescent light detector; and
converting the detected properties into identities of the series of incorporated monomers complementary to a corresponding nucleotide series in the sequence. - View Dependent Claims (43, 44, 45, 46, 47, 48, 49)
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Specification