Methods and Apparatus for Measuring Analytes Using Large Scale FET Arrays
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Abstract
Methods and apparatus relating to very large scale FET arrays for analyte measurements. ChemFET (e.g., ISFET) arrays may be fabricated using conventional CMOS processing techniques based on improved FET pixel and array designs that increase measurement sensitivity and accuracy, and at the same time facilitate significantly small pixel sizes and dense arrays. Improved array control techniques provide for rapid data acquisition from large and dense arrays. Such arrays may be employed to detect a presence and/or concentration changes of various analyte types in a wide variety of chemical and/or biological processes. In one example, chemFET arrays facilitate DNA sequencing techniques based on monitoring changes in hydrogen ion concentration (pH), changes in other analyte concentration, and/or binding events associated with chemical processes relating to DNA synthesis.
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Citations
135 Claims
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1-106. -106. (canceled)
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107. A method for nucleic acid sequencing comprising:
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(a) disposing a plurality of nucleic acid templates at discrete locations on a substrate associated with a semiconductor based sensor array, the discrete locations associated with at least one sensor comprising a field-effect transistor configured to provide output signals representative of a sequencing reaction byproducts proximate thereto; (b) introducing a known nucleotides, primer, and enzyme into the reaction chambers; (c) providing conditions that allow the nucleic acid templates, nucleotide, primer, and enzyme to react; (d) detecting incorporation of the nucleotides by sequencing reaction byproduct accumulation proximate to the at least one sensor; (e) washing unincorporated known nucleotides from the discrete locations; (f) outputting signals from respective sensors in response to detecting the at least one sequencing reaction byproducts; and (g) repeating steps (b) through (d) until a portion of the plurality of nucleic acid templates are at least partially sequenced. - View Dependent Claims (108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130)
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131. A method for sequencing a nucleic acid comprising:
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disposing a plurality of template nucleic acids in a plurality of reaction chambers, wherein the plurality of reaction chambers are disposed proximate to a chemical-sensitive field effect transistor (chemFET) array comprising at least one chemFET for respective reaction chambers, hybridizing the template nucleic acids to primers and subjecting to enzyme activity to synthesize a complementary nucleic acid strand by incorporating one or more known nucleotides sequentially at the 3′
end of the primer; anddetecting incorporation of the one or more known nucleotides by a change in current in the at least one chemFET of the chemFET array. - View Dependent Claims (132)
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133. A method for sequencing a nucleic acid comprising:
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disposing a plurality of template nucleic acids into a plurality of reaction chambers, wherein the plurality of reaction chambers are coupled to a chemical-sensitive field effect transistor (chemFET) array comprising at least one chemFET for respective reaction chambers; incorporating one or more known nucleotides sequentially into newly synthesized nucleic acid strands complementary to the template nucleic acids; and detecting the incorporation of the one or more known nucleotides by detecting the generation of sequencing reaction byproduct by the chemFETs. - View Dependent Claims (134, 135)
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Specification