System for hot-start amplification via a multiple emulsion
First Claim
1. A method of nucleic acid amplification, comprising providing a composition comprising a plurality of compound droplets disposed in an aqueous phase, wherein each compound droplet comprise a plurality of aqueous droplets, each aqueous droplet enclosed by an oil droplet and each aqueous droplet containing at least one amplification reagent;
- heating the composition to cause fusion of the aqueous droplets with the aqueous phase, such that the at least one amplification reagent is added to the aqueous phase to form at least one reaction mixture; and
cycling the at least one reaction mixture thermally to encourage amplification of a nucleic acid target.
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Abstract
System, including methods, apparatus, compositions, and kits, for making and using compound droplets of a multiple emulsion to supply an amplification reagent, such as a heat-stable DNA polymerase or DNA ligase, to an aqueous phase in which the compound droplets are disposed. The compound droplets may be induced to supply the amplification reagent by heating the multiple emulsion, to achieve hot-start amplification.
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Citations
14 Claims
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1. A method of nucleic acid amplification, comprising providing a composition comprising a plurality of compound droplets disposed in an aqueous phase, wherein each compound droplet comprise a plurality of aqueous droplets, each aqueous droplet enclosed by an oil droplet and each aqueous droplet containing at least one amplification reagent;
heating the composition to cause fusion of the aqueous droplets with the aqueous phase, such that the at least one amplification reagent is added to the aqueous phase to form at least one reaction mixture; and
cycling the at least one reaction mixture thermally to encourage amplification of a nucleic acid target.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method of nucleic acid amplification, comprising providing a composition comprising a plurality of compound droplets disposed in an aqueous phase, wherein each compound droplet comprise a plurality of aqueous droplets, each aqueous droplet enclosed by an oil droplet and each aqueous droplet containing a heat stable polymerase;
heating the composition such that the aqueous droplets with the aqueous phase, to form at least one reaction mixture capable of PCR amplification of a nucleic acid target, if present, in the at least one reaction mixture; and
cycling the at least one reaction mixture thermally to encourage PCR amplification.
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12. A composition for nucleic acid amplification, comprising an aqueous phase that is dispersed or continuous;
- a plurality of compound droplet disposed in the aqueous phase;
wherein each compound droplet comprise a plurality of aqueous droplets, each aqueous droplet enclosed by an oil droplet and each aqueous droplet containing at least one amplification reagent, wherein heating the composition causes the aqueous droplets to fuse with the aqueous phase to supply the at least one amplification reagent to the aqueous phase. - View Dependent Claims (13, 14)
- a plurality of compound droplet disposed in the aqueous phase;
Specification