METHODS FOR DETECTING TARGET ANALYTES AND ENZYMATIC REACTIONS
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Abstract
A microsphere-based analytic chemistry system and method for making the same is disclosed in which microspheres or particles carrying bioactive agents may be combined randomly or in ordered fashion and dispersed on a substrate to form an array while maintaining the ability to identify the location of bioactive agents and particles within the array using an optically interrogatable, optical signature encoding scheme. A wide variety of modified substrates may be employed which provide either discrete or non-discrete sites for accommodating the microspheres in either random or patterned distributions. The substrates may be constructed from a variety of materials to form either two-dimensional or three-dimensional configurations. In a preferred embodiment, a modified fiber optic bundle or array is employed as a substrate to produce a high density array. The disclosed system and method have utility for detecting target analytes and screening large libraries of bioactive agents.
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Citations
59 Claims
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1-29. -29. (canceled)
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30. A method of detecting target nucleic acids in a sample, said method comprising:
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(a) providing an array comprising (i) a population of beads comprising a first subpopulation of beads having probe nucleic acids associated therewith and a second subpopulation of beads having enzymes associated therewith, said beads of the first subpopulation being a different size from the beads of the second subpopulation; and (ii) a substrate having a surface comprising a plurality of discrete sites, wherein said population of beads is randomly distributed on said surface such that sites of said plurality of discrete sites have a single bead from the first subpopulation associated therewith, and wherein said sites further comprise beads from said second subpopulation; and (b) providing a sample comprising target nucleic acids; (c) contacting said sample with said array, whereby said target nucleic acids hybridize to said probe nucleic acids; and (d) determining the presence of said target nucleic acids hybridized to said probe nucleic acids. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46)
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47. A method of detecting target nucleic acids in a sample, said method comprising:
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(a) providing a sample comprising target nucleic acids to an array, said array comprising a substrate, said substrate comprising a plurality of wells having a population of beads randomly distributed therein, said population of beads comprising a first subpopulation of beads having probe nucleic acids associated therewith and a second subpopulation of beads having enzymes associated therewith, said beads of the first subpopulation being a different size from the beads of the second subpopulation, and said beads of the first subpopulation being distributed such that wells of said plurality of wells contain a single bead from the first subpopulation of beads; (b) allowing said target nucleic acids to hybridize to said probe nucleic acids; and (c) determining the presence of said target nucleic acids hybridized to said probe nucleic acids. - View Dependent Claims (48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59)
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Specification