Apparatus and methods for parallel processing of micro-volume liquid reactions
First Claim
1. A method for simultaneously conducting a plurality of micro-volume reactions, the method comprising:
- (a) introducing a plurality of liquid samples into the sample chambers of a microhole apparatus, wherein the samples contain necessary reaction components; and
(b) placing the apparatus into an environment favorable to the reaction;
wherein the microhole apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other.
1 Assignment
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Accused Products
Abstract
Disclosed herein are apparatuses and methods for conducting multiple simultaneous micro-volume chemical and biochemical reactions in an array format. In one embodiment, the format comprises an array of microholes in a substrate. Besides serving as an ordered array of sample chambers allowing the performance of multiple parallel reactions, the arrays can be used for reagent storage and transfer, library display, reagent synthesis, assembly of multiple identical reactions, dilution and desalting. Use of the arrays facilitates optical analysis of reactions, and allows optical analysis to be conducted in real time. Included within the invention are kits comprising a microhole apparatus and a reaction component of the method(s) to be carried out in the apparatus.
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Citations
69 Claims
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1. A method for simultaneously conducting a plurality of micro-volume reactions, the method comprising:
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(a) introducing a plurality of liquid samples into the sample chambers of a microhole apparatus, wherein the samples contain necessary reaction components; and
(b) placing the apparatus into an environment favorable to the reaction;
wherein the microhole apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method for adding a component to a micro-volume reaction, wherein the method comprises the steps of:
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(a) providing a first apparatus comprising a first sample chamber containing a reaction mixture;
(b) providing a second apparatus comprising a second sample chamber containing the component; and
(c) bringing the apparatuses into proximity such that liquid contact is established between the first sample chamber and the second sample chamber;
wherein each apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other. - View Dependent Claims (13, 14, 15, 16)
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17. A method for adding a nucleic acid to a micro-volume reaction, wherein the method comprises the steps of:
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(a) providing a first apparatus comprising a first sample chamber containing a reaction mixture;
(b) providing a second apparatus comprising a second sample chamber containing the nucleic acid; and
(c) bringing the apparatuses into proximity such that liquid contact is established between the first sample chamber and the second sample chamber;
wherein each apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber.
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18. A method for introducing a liquid sample into a sample chamber, wherein the method comprises the steps of:
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(a) contacting an apparatus with a liquid solution; and
(b) removing the apparatus from the solution;
wherein the apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other.
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19. A method for introducing a liquid sample comprising a nucleic acid into a sample chamber, wherein the method comprises the steps of:
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(a) contacting an apparatus with a liquid solution comprising a nucleic acid; and
(b) removing the apparatus from the solution;
wherein the apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber.
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20. A method for diluting a solution, wherein the method comprises the steps of:
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(a) providing a first apparatus comprising a first sample chamber containing the solution (b) providing a second apparatus comprising a second sample chamber containing a diluent; and
(c) bringing the apparatuses into proximity such that liquid contact is established between the first sample chamber and the second sample chamber;
wherein each of the apparatuses comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other. - View Dependent Claims (21, 22)
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23. A method for diluting a solution comprising a nucleic acid, wherein the method comprises the steps of:
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(a) providing a first apparatus comprising a first sample chamber containing the solution which comprises a nucleic acid;
(b) providing a second apparatus comprising a second sample chamber containing a diluent; and
(c) bringing the apparatuses into proximity such that liquid contact is established between the first sample chamber and the second sample chamber;
wherein each of the apparatuses comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber. - View Dependent Claims (24, 25)
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26. A method for selective retention of a molecule in a first sample chamber, wherein the method comprises the steps of:
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(a) providing a first apparatus, wherein the first sample chamber contains a solution comprising the molecule and one or more additional solute molecules of higher diffusibility;
(b) providing a second apparatus comprising a second sample chamber containing a solvent;
(c) bringing the apparatuses into proximity such that liquid contact is established between the first sample chamber and the second sample chamber; and
(d) removing the apparatuses from proximity;
wherein each of the apparatuses comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other. - View Dependent Claims (27, 28, 29, 30)
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31. A method for selective retention of a nucleic acid in a first sample chamber, wherein the method comprises the steps of:
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(a) providing a first apparatus, wherein the first sample chamber contains a solution comprising the nucleic acid and one or more additional solute molecules of higher diffusibility;
(b) providing a second apparatus comprising a second sample chamber containing a solvent;
(c) bringing the apparatuses into proximity such that liquid contact is established between the first sample chamber and the second sample chamber; and
(d) removing the apparatuses from proximity;
wherein each of the apparatuses comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber. - View Dependent Claims (32, 33, 34, 35)
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36. A method for parallel electrophoretic analysis of a plurality of micro-volume reactions, wherein the method comprises:
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(a) conducting the reactions in a microhole apparatus;
(b) placing the apparatus in contact with an electrophoresis medium; and
(c) conducting electrophoresis;
wherein the apparatus comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber. - View Dependent Claims (37, 38, 39)
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40. A method for preparing a plurality of samples for mass spectrometric analysis, wherein the samples are placed in an apparatus that comprises a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber:
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(a) extends through the substrate;
(b) comprises one or more walls and an opening at each end; and
(c) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber. - View Dependent Claims (41, 42, 43)
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44. A method for mixing a plurality of micro-volume samples, the method comprising:
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(a) providing a first microhole apparatus comprising a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber;
(b) providing a second microhole apparatus comprising a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber;
(i) extends through the substrate;
(ii) comprises one or more walls and an opening at each end; and
(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
(c) bringing the apparatuses into proximity such that liquid contact is established between more than one sample chamber from the first apparatus and a sample chamber in the second apparatus.
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45. An apparatus for containing multiple micro-volume liquid samples comprising a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber:
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(a) extends through the substrate, (b) comprises one or more walls and an opening at each end, and (c) holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other. - View Dependent Claims (46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62)
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63. A kit comprising an apparatus for containing multiple micro-volume liquid samples comprising a substrate, wherein the substrate defines a plurality of sample chambers, wherein each sample chamber:
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(a) extends through the substrate, (b) comprises one or more walls and an opening at each end, and (c) holds a sample such that the sample is in the form of a thin film such that a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber; and
wherein the sample chamber has a height to width ratio of less than 1;
1, wherein the height of the sample chamber is measured from one face of the substrate to the other; and
further comprising a reaction component packaged in a suitable container. - View Dependent Claims (64, 65, 66, 67, 68, 69)
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Specification