Method and Apparatus for Rapid Nucleic Acid Sequencing
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Abstract
Methods and apparatus relating to FET arrays including large FET arrays for monitoring chemical and/or biological reactions such as nucleic acid sequencing-by-synthesis reactions. Some methods provided herein relate to improving signal (and also signal to noise ratio) from released hydrogen ions during nucleic acid sequencing reactions.
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Citations
183 Claims
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1-154. -154. (canceled)
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155. A method for nucleic acid sequencing, comprising:
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a) preparing a nicked nucleic acid; b) providing a sequencing reaction mixture comprising the nicked nucleic acid, a known nucleotide, a nucleic acid polymerase with exonuclease activity in a reaction chamber, wherein the reaction chamber is coupled to a field effect transistor (FET); c) detecting a signal in the FET in response to excision of a first nucleotide at a first position on the nicked nucleic acid and incorporation of the known nucleotide at a second position on the nicked nucleic acid, thereby determining the identity of a nucleotide complementary to the incorporated known nucleotide in the nicked nucleic acid; and d) repeating steps b) and c) with additional known nucleotides to determine the identity of a plurality of nucleotides in the nicked nucleic acid. - View Dependent Claims (156, 157, 158, 159, 160, 161, 162)
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163. A method for sequencing a plurality of nucleic acids, comprising:
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a) providing a plurality of nicked nucleic acids disposed in an array of reaction chambers, wherein the reaction chambers are associated with at least one field effect transistor (FET); b) contacting the nicked nucleic acids with a known nucleotide and an enzymatic composition having polymerase and exonuclease activity; c) detecting signals using the at least one FET in response to excisions and incorporations in the plurality of nicked nucleic acids; d) determining the identity of nucleotides within the nicked nucleic acids on the basis of the detected signals and the identity of the known nucleotide; and e) repeating steps b) and d) with additional known nucleotides to determine at least a portion of the sequence for the plurality of nicked nucleic acids. - View Dependent Claims (164, 165, 166, 167, 168, 169, 170, 171, 172)
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173. A method for sequencing a plurality of nucleic acids, comprising:
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a) providing a plurality of nicked nucleic acids disposed in an array of reaction chambers, wherein the reaction chambers are associated with at least one field effect transistor (FET); b) contacting the nicked nucleic acids with a known nucleotide and an enzymatic composition having polymerase and exonuclease activity; c) detecting a signal in the at least one FET in response to excision of a first nucleotide at a first position in the plurality of nicked nucleic acids and incorporation of the known nucleotide at a second position in the plurality of nicked nucleic acids; d) determining the identity of nucleotides complementary to the incorporated nucleotides within the nicked nucleic acids on the basis of the detected signals and the identity of the known nucleotide; and e) repeating steps b) and d) with additional known nucleotides to determine at least a portion of the sequence for the plurality of nicked nucleic acids. - View Dependent Claims (174)
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- 175. A composition comprising a nicked nucleic acid, a nucleotide, and one or more polymerases disposed in a reaction chamber capacitively coupled to a field effect transistor (FET).
Specification