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US 2019/0292582 A1Published application
Analyte Enrichment Methods and Compositions
AssigneeQiagen Sciences, LLC
InventorsJerzy Olejnik, Steven Gordon, Martina Werner
Life of this patent
Estimating the term…
Abstract
Methods and compositions for enriching a population of particles containing an analyte are disclosed. In one embodiment, enrichment beads are used that are larger in size than the beads used for amplification. A separation device is employed that can retain larger beads with bound amplified beads. The technique finds many uses, including enriching for beads with clonally amplified template, which can be used in a variety of assays, including nucleic acid sequencing.
Claims28
1-44. (canceled)
A method of enriching, comprising:
- a) providing
- i) an emulsion comprising one or more aqueous compartments in oil, at least some of said compartments comprising PCR reagents, a first PCR primer immobilized on an emulsion bead, a second biotinylated PCR primer in solution, and a nucleotide sequence template;
- and
- ii) enrichment beads, wherein said enrichment beads are larger than emulsion beads in said compartments;
- b) exposing said emulsion to conditions so as to amplify at least some of said template to produce amplified template on at least some of said emulsion beads in at least some of said compartments, wherein one strand of said amplified template terminates with biotin;
- and
- c) enriching for emulsion beads comprising said amplified template by contacting said emulsion beads with said enrichment beads, wherein said emulsion beads comprising said amplified template bind to said enrichment beads so as to make a population of emulsion bead—enrichment bead complexes;
- and
- d) capturing at least some of said population of complexes under conditions such that a majority of said emulsion beads not comprising amplified template are not captured.
6 more elements
Dependencies are inferred from the claim text.