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US 12,435,367 B2Granted
Apparatus, system, and method using immiscible-fluid-discrete-volumes
AssigneeApplied Biosystems, LLC
InventorsLinda G. Lee, Sam L Woo, Congcong Ma, Richard T. Reel, Mark F. Oldham, David M. Cox, Benjamin G. Schroeder, Jon M. Sorenson, Willy Wiyatno
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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.
Claims20
A method comprising:
- contacting a stream of aqueous sample fluid flowing in a first conduit with a stream of non-aqueous spacing fluid that is immiscible with the aqueous sample fluid to form discrete volumes of the aqueous sample fluid separated from one another by the non-aqueous spacing fluid, wherein the aqueous sample fluid comprises target nucleic acid, and wherein a first plurality of the discrete volumes contains at least one molecule comprising the target nucleic acid and a second plurality of the discrete volumes contains no molecules comprising the target nucleic acid;
- amplifying the target nucleic acid in one or more of the first plurality of the discrete volumes to form an amplicon;
- after amplifying the target nucleic acid, adding additional spacing fluid between consecutive discrete volumes of one or both of the first plurality of discrete volumes and the second plurality of discrete volumes, so as to adjust or control spacing between the consecutive discrete volumes;
- in a second conduit, detecting a fluorescence signal from the amplicon in the one or more of the first plurality of the discrete volumes;
- and
- based on the detecting, discriminating between the one or more of the first plurality of the discrete volumes and the second plurality of the discrete volumes.
3 more elements
A method comprising:
- forming droplets of an aqueous sample fluid separated from one another by a spacing fluid that is immiscible with the aqueous sample fluid, the aqueous sample fluid comprising a polynucleotide comprising a target nucleic acid sequence, wherein a first plurality of droplets contains at least one molecule of the polynucleotide comprising the target nucleic acid sequence per droplet and a second plurality of droplets contains no molecules of the polynucleotide comprising the target nucleic acid sequence;
- performing an amplification assay on the polynucleotide in one or more of the first plurality of droplets to form an amplicon;
- after performing the amplification assay, adding additional spacing fluid between consecutive aqueous droplets of one or both of the first plurality of aqueous droplets and the second plurality of aqueous droplets, so as to adjust or control the spacing between the consecutive aqueous droplets;
- and
- monitoring for a fluorescence signal from one or more of the first plurality of droplets, the fluorescent signal indicative of a presence of the amplicon.
2 more elements
Dependencies are inferred from the claim text.