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 the concentration of inorganic pyrophosphate (PPi), hydrogen ions, and nucleotide triphosphates.
229 Citations
77 Claims
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1-8. -8. (canceled)
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9. A method for sequencing a nucleic acid comprising:
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contacting a template nucleic acid with a sequencing primer and a polymerase for times and conditions sufficient to allow the template nucleic acid to bind to the sequencing primer for form a template/primer hybrid and to allow the polymerase to bind to the template/primer hybrid, and synthesizing a new nucleic acid strand by incorporating one or more known nucleotide triphosphates sequentially at the 3′
end of the sequencing primer in a low ionic strength environment, anddetecting incorporation of the one or more known nucleotide triphosphates by detection of one or more corresponding ionic pulses with a chemFET. - View Dependent Claims (32, 34)
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10-22. -22. (canceled)
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23. A method for sequencing a nucleic acid comprising:
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(a) disposing a plurality of beads into a plurality of reaction chambers, each reaction chamber comprising a single bead, each bead attached to a plurality of identical template nucleic acids, each of the template nucleic acids hybridized to a sequencing primer and bound to a polymerase, and each reaction chamber in contact with or capacitively coupled to at least one chemical-sensitive field effect transistor (chemFET), (b) introducing a plurality of known identical nucleotide triphosphates into each reaction chamber, (c) detecting sequential incorporation at the 3′
end of the sequencing primer of one or more nucleotide triphosphates if complementary to corresponding nucleotides in the template nucleic acid,(d) washing unincorporated nucleotide triphosphates from the reaction chambers, and (e) repeating steps (b) through (d) in the same reaction chamber using a different plurality of known nucleotide triphosphates. - View Dependent Claims (33, 41, 44, 49, 52, 59, 60, 61, 62, 64, 65, 70, 75, 76)
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24-30. -30. (canceled)
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31. A method for sequencing a nucleic acid comprising:
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disposing a plurality of identical template nucleic acids into a reaction chamber, wherein the reaction chamber is in contact with or capacitively coupled to a chemical-sensitive field effect transistor (chemFET), and wherein each of the template nucleic acids is hybridized to a sequencing primer and is bound to a polymerase, incorporating one or more known nucleotide triphosphates sequentially at the 3′
end of the sequencing primer, anddetecting the incorporation of the one or more known nucleotide triphosphates by an electrical pulse at the chemFET, wherein the incorporation of 10-1000 nucleotide triphosphates is detected. - View Dependent Claims (35)
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36-40. -40. (canceled)
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42-43. -43. (canceled)
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45-48. -48. (canceled)
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50-51. -51. (canceled)
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53-58. -58. (canceled)
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63. (canceled)
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66-69. -69. (canceled)
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71-74. -74. (canceled)
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77-246. -246. (canceled)
Specification