Target capture and barcoding in monodisperse droplets
AssigneeFluent Biosciences Inc.
InventorsRobert Meltzer, Kristina Fontanez, Desiree Schenk, Yi Xue
Life of this patent
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Abstract
The methods and systems described herein provide capture template particles and an improved emulsion droplet-based target capture and barcoding method thereof. The capture template particles and methods disclosed herein allow capturing targets of interest from biological samples, and barcoding of specific nucleic acids contained in the captured targets. The nucleic acids can be contained within living or nonliving structures, including particles, viruses, and cells. The nucleic acids can include, e.g., DNA or RNA.
Claims21
A capture template particle comprising:
- a template particle comprising one or more tethering moieties, wherein each of the one or more tethering moieties is attachable to one or more target-specific elements.
A method of target capture and barcoding in monodisperse droplets comprising:
- combining a plurality of capture template particles with a first fluid to provide a first mixture, wherein the first fluid comprises a plurality of target particles;
- incubating the first mixture to allow association of the plurality of target particles to the plurality of capture template particles, thereby a portion of the plurality of target particles become associated to the capture template particles;
- generating a supernatant and pellet from the first mixture, optionally by centrifuging;
- removing the supernatant to separate the plurality of capture template particles and associated target particles from the solution of the first mixture;
- washing and resuspending the pellet in a reaction buffer to provide a second mixture;
- combining the second mixture with a second fluid to provide a third mixture, wherein the second fluid is immiscible with the second mixture;
- and
- shearing the third mixture such that a plurality of the capture template particles are encapsulated in a plurality of monodisperse droplets in the second fluid, thereby providing a plurality of monodisperse droplets comprising reaction buffer, one of the capture template particles, and one of the plurality of target particles associated to said capture template particles.
21-95. (canceled)
Dependencies are inferred from the claim text.