Analysis devices, kits, and related methods for digital quantification of nucleic acids and other analytes
First Claim
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1. A method of amplifying a nucleic acid molecule, comprising:
- (a) introducing a sample comprising at least one nucleic acid molecule within a plurality of first areas in a first substrate and a plurality of first areas in a second substrate, wherein said first and second substrates are configured in a first position with the plurality of first areas in said first substrate overlapping with the plurality of first areas in said second substrate to form a continuous fluidic path within said first and second substrates;
(b) contacting the sample disposed in the plurality of first areas in the first substrate with at least one component of an amplification reagent disposed in a plurality of second areas in the second substrate,the contacting being effected by relative motion between said first substrate and said second substrate to a second position, wherein said plurality of first areas in said first substrate are isolated from the plurality of first areas in said second substrate, thereby placing at least some of the first areas in said first substrate into direct fluid communication with at least some of the plurality of second areas so as to form a plurality of closed reaction chambers; and
(c) exposing the plurality of closed reaction chambers having the at least one nucleic acid molecule to conditions effective for amplification of the at least one nucleic acid molecule.
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Abstract
Provided are devices and methods for effecting processing of samples, including essentially isothermal amplification of nucleic acids, at multiple reaction locations in a single device. In some embodiments, the disclosed devices and methods provide for effecting parallel sample processing in several hundred locations on a single device.
258 Citations
15 Claims
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1. A method of amplifying a nucleic acid molecule, comprising:
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(a) introducing a sample comprising at least one nucleic acid molecule within a plurality of first areas in a first substrate and a plurality of first areas in a second substrate, wherein said first and second substrates are configured in a first position with the plurality of first areas in said first substrate overlapping with the plurality of first areas in said second substrate to form a continuous fluidic path within said first and second substrates; (b) contacting the sample disposed in the plurality of first areas in the first substrate with at least one component of an amplification reagent disposed in a plurality of second areas in the second substrate, the contacting being effected by relative motion between said first substrate and said second substrate to a second position, wherein said plurality of first areas in said first substrate are isolated from the plurality of first areas in said second substrate, thereby placing at least some of the first areas in said first substrate into direct fluid communication with at least some of the plurality of second areas so as to form a plurality of closed reaction chambers; and (c) exposing the plurality of closed reaction chambers having the at least one nucleic acid molecule to conditions effective for amplification of the at least one nucleic acid molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method of effecting amplification of at least one nucleic acid target molecule, comprising:
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(a) introducing a sample material comprising a nucleic acid target within a plurality of first areas in a first substrate and a plurality of first areas in a second substrate, wherein said first and second substrates are configured in a first position with the plurality of first areas in said first substrate overlapping with the plurality of first areas in said second substrate to form a continuous fluidic path within said first and second substrates; (b) contacting (1) the sample material disposed in the plurality of first areas in the first substrate, at least one of the first areas in the first substrate containing one molecule of the nucleic acid target, with (2) a reactant material disposed in a plurality of second areas in the second substrate, the contacting being effected by relative motion between said first substrate and said second substrate to a second position, wherein said plurality of first areas in said first substrate are isolated from the plurality of first areas in said second substrate, resulting in pairwise placement of at least some of the first areas and at least some of the second areas into direct fluid communication with one another so as to form a plurality of closed reaction chambers, the contacting effecting amplification of at least one nucleic acid target molecule. - View Dependent Claims (14, 15)
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Specification