Method for conducting binding reactions on a solid surface within an enclosed chamber
First Claim
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1. A method, comprising:
- (a) providing a device comprised of a (i) a substrate having a surface with at least a portion of said surface comprising a molecular probe, and (ii) a cover which sealingly contacts the substrate surface, to form an enclosure having an interior space comprising a sealed hybridization chamber having a height ranging from about 50 μ
m to about 500 μ
m; and
(b) introducing into the chamber, a sample fluid comprising (i) a target molecule which may bind to a surface-bound molecular probe, (ii) a hybridization buffer, (iii) a surfactant of a type and present at a concentration effective to substantially reduce nonspecific binding and promote mixing of components within the sample fluid and (iv) a bubble; and
(c) mixing the sample fluid by moving the chamber to cause the bubble to displace the sample fluid and maintaining conditions within the chamber for a period of time sufficient to allow binding between the target molecule and a surface-bound molecular probe to occur.
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Abstract
The invention provides an apparatus and method for conducting chemical or biochemical reactions on a solid surface within an enclosed chamber. The invention may be used in conducting hybridization reactions, as of biopolymers such as DNA, RNA, oligonucleotides, peptides, polypeptides, proteins, antibodies, and the like. In another aspect, the invention provides an improved method for mixing a thin film of solution, as in a hybridization chamber. The invention further provides a kit for carrying out the methods of the invention.
51 Citations
22 Claims
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1. A method, comprising:
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(a) providing a device comprised of a (i) a substrate having a surface with at least a portion of said surface comprising a molecular probe, and (ii) a cover which sealingly contacts the substrate surface, to form an enclosure having an interior space comprising a sealed hybridization chamber having a height ranging from about 50 μ
m to about 500 μ
m; and(b) introducing into the chamber, a sample fluid comprising (i) a target molecule which may bind to a surface-bound molecular probe, (ii) a hybridization buffer, (iii) a surfactant of a type and present at a concentration effective to substantially reduce nonspecific binding and promote mixing of components within the sample fluid and (iv) a bubble; and (c) mixing the sample fluid by moving the chamber to cause the bubble to displace the sample fluid and maintaining conditions within the chamber for a period of time sufficient to allow binding between the target molecule and a surface-bound molecular probe to occur. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A method comprising:
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(a) sealingly contacting a cover to a first substrate having a molecular probe bound to the surface of the first substrate to form a first sealed chamber having a height ranging from about 50 μ
m to about 500 μ
m about the substrate surface-bound molecular probe,(b) performing a binding assay within the first sealed chamber and a sample comprising a target molecule which may bind to the surface-bound molecular probe, (c) opening the chamber and removing the first substrate, (d) reusing the cover by sealingly contacting the cover to a second substrate having a molecular probe bound to the surface of the second substrate, wherein the cover and substrate surface form a second sealed chamber about the substrate surface-bound molecular probe, (e) performing a binding assay with the second sealed chamber and a sample comprising a target molecule which may bind to a surface-bound molecular probes, wherein at least one of (i) the first substrate and the cover and (ii) the second substrate and the cover is compressed together by tightening screws during a step of sealingly contacting. - View Dependent Claims (22)
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Specification