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.
2 Citations
45 Claims
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1-25. -25. (canceled)
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26. A method of making an array, said method comprising:
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(a) providing a substrate with a surface comprising a plurality of discrete sites wherein said substrate is not a fiber optic bundle; (b) providing a population of beads comprising a first subpopulation of beads having 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 (c) randomly distributing said population of beads on said surface such that sites of said plurality of discrete site have a single bead from said first subpopulation associated therewith and such that beads from said second subpopulation end up at said sites. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34)
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35. A method of making an array, said method comprising:
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(a) providing a substrate comprising a plurality of wells wherein said substrate is not a fiber optic bundle; (b) randomly distributing beads of a first subpopulation of beads into wells of said plurality of wells such that the wells contain a single bead from the first subpopulation of beads, wherein beads of the first subpopulation of beads have nucleic acids associated therewith; and (c) distributing beads of a second subpopulation of beads into the wells containing a single bead from the first subpopulation of beads, wherein beads of the second subpopulation of beads have enzymes associated therewith, and wherein beads of the second subpopulation of beads are smaller than beads of said first population of beads. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42)
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43. A method of detecting a nucleic acid on an array, said method comprising:
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(a) providing a substrate with a surface comprising a plurality of discrete sites wherein said substrate is not a fiber optic bundle; (b) providing a population of beads comprising a first subpopulation of beads having 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; (c) randomly distributing said population of beads on said surface such that sites of said plurality of discrete site have a single bead from said first subpopulation associated therewith and such that beads from said second subpopulation end up at said sites; (d) contacting said substrate with a sample comprising target nucleic acids, whereby said target nucleic acids hybridize to the nucleic acids associated with said first population of beads; and (e) determining the presence of said target nucleic acids hybridized to said probe nucleic acids.
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44. A method of detecting a nucleic acid on an array, said method comprising:
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(a) providing a substrate comprising a plurality of wells wherein said substrate is not a fiber optic bundle; (b) randomly distributing beads of a first subpopulation of beads into wells of said plurality of wells such that the wells contain a single bead from the first subpopulation of beads, wherein beads of the first subpopulation of beads have nucleic acids associated therewith; and (c) distributing beads of a second subpopulation of beads into the wells containing a single bead from the first subpopulation of beads, wherein beads of the second subpopulation of beads have enzymes associated therewith, and wherein beads of the second subpopulation of beads are smaller than beads of said first population of beads (d) contacting said substrate with a sample comprising target nucleic acids, whereby said target nucleic acids hybridize to the nucleic acids associated with said first population of beads; and (e) determining the presence of said target nucleic acids hybridized to said probe nucleic acids. - View Dependent Claims (45)
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Specification