Apparatus, System, And Method Using Immiscible-Fluid-Discrete-Volumes
First Claim
Patent Images
1. A method comprising:
- amplifying a nucleic acid in at least one conduit to form an amplicon, the at least one conduit comprising an inner wall;
attaching the amplicon to the inner wall to form an attached amplicon; and
detecting the attached amplicon or an attached derivative thereof, in the at least one conduit.
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Accused Products
Abstract
Various embodiments of the teachings relate to a system or method for sample preparation or analysis in biochemical or molecular biology procedures. The sample preparation can involve small volume processed in discrete portions or segments or slugs, herein referred to as discrete volumes. A molecular biology procedure can be nucleic acid analysis. Nucleic acid analysis can be an integrated DNA amplification/DNA sequencing procedure.
43 Citations
50 Claims
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1. A method comprising:
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amplifying a nucleic acid in at least one conduit to form an amplicon, the at least one conduit comprising an inner wall; attaching the amplicon to the inner wall to form an attached amplicon; and detecting the attached amplicon or an attached derivative thereof, in the at least one conduit.
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2. A method comprising:
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sequentially contacting an aqueous sample fluid in a conduit with a non-aqueous spacing fluid that is immiscible with the aqueous sample, to form a plurality of discrete volumes of the aqueous sample fluid separated from one another by the non-aqueous spacing fluid, the aqueous sample fluid comprising a plurality of target nucleic acid sequences, wherein at least one of the discrete volumes contains at least one target nucleic acid sequence; amplifying the at least one target nucleic acid in the conduit to form an amplicon; and subjecting the amplicon to a nucleic acid sequencing reaction in the conduit. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method comprising:
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flowing a first fluid and a second fluid into a conduit, the first fluid and the second fluid being immiscible with respect to one another, the first fluid containing a plurality of target molecules, wherein the maximum cross-sectional dimension of the conduit is such that a plurality of discrete volumes of the first fluid are formed in the conduit and separated from one another by the second fluid and at least one of the discrete volumes of the first fluid contains a single target molecule. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20)
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21. A system comprising:
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at least one conduit comprising an inner surface and having a maximum cross-sectional dimension; an aqueous sample fluid introduction unit in fluid communication with the at least one conduit; a spacing fluid introduction unit in fluid communication with the at least one conduit, the aqueous sample fluid introduction unit and the spacing fluid introduction unit comprising separate units each in fluid communication with the at least one conduit; and a control unit adapted to flow an aqueous sample fluid and a spacing fluid from the aqueous sample fluid introduction unit and the spacing fluid introduction unit, respectively, and adapted to inject volumes of aqueous sample fluid and spacing fluid that respectively form discrete volumes in the at least one conduit wherein each discrete volume comprises a dimension that is at least about 95% of the maximum cross-sectional dimension. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28)
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29. A system comprising:
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a plurality of sample discrete volumes in a conduit, the discrete volumes being spaced-apart from one another by a spacing fluid that is immiscible with respect to the discrete volumes; and at least one fiducial marker, the at least one fiducial marker identifying at least one volume of the discrete volumes. - View Dependent Claims (30, 31, 32, 33)
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34. A system comprising a first set of discrete volumes and a second set of discrete volumes, in a conduit, the discrete volumes of each set being spaced-apart from one another by a spacing fluid that is immiscible with respect to the discrete volumes;
- wherein the discrete volumes of the first set each have a first length, the discrete volumes of the second set each have a second length, and the first length and second length are different from one another.
- View Dependent Claims (35)
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36. A method comprising:
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generating, in a conduit, a plurality of discrete volumes, spaced apart from one another by a spacing fluid that is immiscible with the discrete volumes, at least one of the discrete volumes containing a single target nucleic acid sequence, the target nucleic acid sequence having a first universal sequence of a universal sequence pair ligated to one end thereof and the second universal sequence of a universal sequence pair ligated to the other end thereof; and reacting the target nucleic acid sequence with the universal primers corresponding to the universal sequence pair, in the respective discrete volume. - View Dependent Claims (37, 38, 39)
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40. A method comprising:
generating, in a conduit, a plurality of discrete volumes, spaced apart from one another by a spacing fluid that is immiscible with the discrete volumes, at least one of the discrete volumes containing a plurality of target nucleic acid sequences and a plurality of pairs of tailed sequence-specific primers. - View Dependent Claims (41, 42)
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43. A nucleic acid identifying system comprising:
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an identification sequence-tagged tailed forward primer comprising a first identification sequence tag, a forward universal primer sequence, and a sequence-specific forward primer; and a tailed reverse primer comprising, a reverse universal primer sequence, and a sequence specific reverse primer; wherein the sequence-specific forward primer and the sequence specific reverse primer are adapted to participate in a nucleic acid amplification reaction of a specific nucleic acid sequence. - View Dependent Claims (44, 45, 46, 47)
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48. A method of nucleic acid sample tracking, comprising:
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providing a tailed forward primer comprising a 5′
end, an identification sequence comprising a 3′
end, and a splint;annealing the splint to the tailed forward primer and the identification sequence such that the end of the tailed forward primer and the end of the identification nucleic acid sequence are disposed adjacent to each other; and ligating the tailed forward primer to the identification sequence, thereby forming an identification sequence tailed forward primer. - View Dependent Claims (49, 50)
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Specification