CLAIMS
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`WHATIS CLAIMED IS:
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`1.
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`a.
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`A system for detecting droplets, comprising:
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`a detector device comprising an input flow path, an intersection region, and an output
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`flow path, wherein the intersection region is downstream ofthe input flow path and the output
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`flow path is downstream ofthe intersection region;
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`b.
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`c.
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`droplets located within the input flow path; and
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`an aqueousfluid for separating the droplets wherein the droplets are introduced to the
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`aqueousfluid at the intersection region.
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`2.
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`The system of claim 1, wherein the input flow path comprises a continuous phase of non-
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`aqueousfluid .
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`3.
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`4.
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`5.
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`The system of claim 2, wherein the non-aqueousfluid is an aqueous-immiscible fluid.
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`The system of claim 2, wherein the non-aqueousfluid is an oil.
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`The system of claim 1, wherein the output flow path comprises a continuous phase of
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`aqueousfluid .
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`6.
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`7.
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`The system of claim 5, wherein the aqueousfluid comprises a surfactant.
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`The system of claim 1, wherein the droplets in the output flow path have an inner core
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`containing an aqueousfluid that is encapsulated with a non-aqueousfluid.
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`8.
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`9.
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`10.
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`a.
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`The system of claim 7, wherein the non-aqueousfluid is a continuous phase.
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`The system of claim 7, wherein the non-aqueousfluid is a discontinuous phase.
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`A-system for detecting droplets, comprising:
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`a detector device comprising an input flow path, an intersection region, and an output
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`flow path, wherein the intersection region is downstream ofthe input flow path and the output
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`flow path is downstream ofthe intersection region; and
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`b.
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`an oil-immiscible fluid for separating the droplets, wherein the oil-immiscible fluid is
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`introducedto the droplets at the intersection region;
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`wherein the continuous phase offluid within the input flow path is a non-aqueousfluid and
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`wherein the inner surface of the output flow path is coated with the non-aqueousfluid with a
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`thicknessthatis at least 0.01% of the diameter of the outflow path, thereby narrowing the
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`aperture of the output flow path; and
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`wherein the droplets in the output flow path have an inner core containing an aqueousfluid that
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`is encapsulated with the non-aqueousfluid.
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`11.
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`The system of claim 10, wherein the thicknessis at least 0.1% of the diameter of the
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`output flow path.
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`12.|The system of claim 10, wherein the thicknessis at least 1% of the diameter of the output
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`flow path.
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`13.
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`The system of claim 10, wherein the thicknessis at least 5% of the diameter of the output
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`flow path.
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`14.
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`The system of claim 10, wherein the thicknessis in a range of 1%-90% of the diameter of
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`the output flow path.
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`15.|The system of claim 1 or 10, wherein the droplets flow through the output flow path
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`serially and substantially centered .
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`16.
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`17.
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`The system of claim 1 or 10, wherein the droplets comprise a nucleic acid.
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`The system of claim 1 or 10, wherein the droplets comprise a product of a nucleic acid
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`amplification reaction.
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`18.|The system of claim 1 or 10, wherein the droplets are emulsified droplets.
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`19.
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`The system of claim 1 or 10, wherein each of the droplets comprises, on average, less
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`than five target nucleic acids.
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`20.
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`21.
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`22.
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`23.
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`The system of claim 10, wherein the oil-immiscible fluid comprises a surfactant.
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`The system of claim 6 or 20, wherein the surfactant is ionic surfactant.
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`The system of claim 6 or 20, wherein the surfactant is non-ionic surfactant.
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`The system of claim 6 or 20, wherein the surfactant is greater than 0.01 % of the weight
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`of the total aqueousfluid.
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`24.
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`The system of claim 6 or 20, wherein the surfactant is greater than 0.01 % of the weight
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`of the total aqueousfluid.
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`25.
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`The system of claim 6 or 20, wherein the surfactant is greater than 0.1 % of the weight of
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`the total aqueous fluid.
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`26.
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`The system of claim 6 or 20, wherein the surfactant is greater than 0.5 % of the weight of
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`the total aqueous fluid.
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`27.
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`The system of claim 6 or 20, wherein the surfactant is in a range of 0.5% to 95.0% ofthe
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`weight of the total aqueousfluid, inclusive.
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`28.|Amethodof separating droplets, comprising:
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`(a) flowing a stream of non-aqueousfluid comprising the droplets along a flow path comprising:
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`(1) an input flow path,(11) an intersection region, and (iii) a downstream output flow path; and
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`(b) introducing a stream of oil-immiscible fluid to the intersection region;
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`wherein the average distance between the droplets in the output flow path is greater than the
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`average distance between the droplets within the input flow path.
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`29.
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`A methodof separating droplets, comprising:
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`(a) flowing a stream of non-aqueousfluid comprising the droplets along a flow path comprising:
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`(1) an intersection region and(ii) a downstream output flow path; and
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`(b) introducing a stream of oil-immiscible fluid to the intersection region;
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`wherein the droplets are heated prior to entering the intersection region.
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`30.
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`A methodof detecting droplets, comprising:
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`(a) flowing a stream of non-aqueousfluid through a continuous flow path comprising an
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`intersection region and a downstream detection region, wherein the non-aqueousfluid comprises
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`the droplets;
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`(b) introducing a stream of oil-immiscible fluid to the intersection region; and
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`(c) detecting a signal from the droplets as they pass through the downstream detection region.
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`31.
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`The method of claim 28 or 29, wherein the output flow path comprises: (a) a continuous
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`phaseofoil-immiscible fluid; and (b) aqueous droplets encapsulated by a layer of non-aqueous
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`fluid.
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`32.
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`The method of any one of claims 28-30, wherein the flow paths of the non-aqueousfluid
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`and that of the oil-immiscible fluid are substantially perpendicular.
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`33.
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`34.
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`35.
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`|The method ofany one of claims 28-30, wherein the oil-immiscible fluid comprises water.
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`The method of any one of claims 28-30, wherein the oil-immiscible fluid comprises air.
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`The methodof claim 33, wherein the water comprises a surfactant.
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`36.|The methodof claim 35, wherein the weight of the surfactant is at least 0.001% ofthe
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`weight of the water.
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`37.|The methodof claim 35, wherein the weight of the surfactantis at least 0.01% of the
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`weight of the water.
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`38.
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`The methodof claim 35, wherein the weight of the surfactantis at least 0.1% of the
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`weight of the water.
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`39.
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`The method of claim 35, wherein the weight of the surfactantis at least 1% of the weight
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`of the water.
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`40.
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`The method ofclaim 35, wherein the weight of the surfactant is in a range of 0.1%-99%
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`of the weight of the water.
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`41.
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`42.
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`The method of claim 33, wherein the water comprises glycerol.
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`The methodof claim 41, wherein the weight of the glycerolis at least 0.01 % of the
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`weight of the water.
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`43.|The method of claim 41, wherein the weight of the glycerol is at least 0.1 % of the weight
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`of the water.
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`44.
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`The method ofclaim 41, wherein the weight of the glycerolis in a range of 0.1 % ofthe
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`weight of the water.
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`45.
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`The method ofclaim 33, wherein the water comprises an antimicrobial agent.
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`46.|The method ofclaim 33, wherein the water comprises an antifoaming agent.
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`47.
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`The method ofany one of claims 28-30, wherein the non-aqueousfluid comprises an oil
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`selected from the group consisting of a silicone oil, a mineral oil, a hydrocarbonoil, a
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`fluorocarbon oil, a vegetable and a soybeanoil.
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`48.
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`49.
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`The method of claim 47, wherein the oil comprises a surfactant .
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`The method ofclaim 48, wherein the surfactant is selected from the group consisting of a
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`fluorocarbon, a hydrocarbonora silicone surfactant.
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`50.|The methodof claim 48, wherein the surfactant comprises a fluorinated surfactant.
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`51.
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`The method of claim 50, wherein the fluorinated surfactant is negatively charged.
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`52.|The methodof claim 50, wherein the fluorinated surfactant comprises a carboxylate
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`group.
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`53.|The method of any one of claims 28-30, wherein the droplets comprise aqueous droplets
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`encapsulated by the non-aqueousfluid.
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`54.|The method of claim 53, wherein the aqueous phase of the droplets comprises a
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`surfactant.
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`55.
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`56.
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`57.
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`The method of claim 54, wherein the surfactant is ionic surfactant.
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`The method of claim 54, wherein the surfactant is non-ionic surfactant.
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`The methodof claim 54, wherein the surfactant is a block copolymer of polypropylene
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`oxide and polyethylene oxide.
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`58.
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`The methodof claim 28 or 29, wherein the average distance between the droplets in the
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`output flow pathis at least 1.2 times the average distance between the droplets in the input flow
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`path.
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`59.|The method of any one of claims 28-30, wherein the step of flowing the droplets
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`comprises a step of operating one or more syringe pumps.
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`60.|The method of claim 28 or 29, wherein the droplets are substantially centered within the
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`output flow path.
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`61.
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`The method of any one of claims 28-30, wherein the oil-immiscible fluid formsa virtual
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`capillary within the output flow path.
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`62.|The method of any one of claims 28-30, wherein the droplets have varyingsizes.
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`63.|The method of any one of claims 28-30, wherein the droplets are emulsified droplets.
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`64.
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`The method of any one of claims 28-30, wherein the droplets comprise a nucleic acid.
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`65.|The method of any one of claims 28-30, wherein the droplets comprise a productof a
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`nucleic acid amplification reaction.
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`66.|The method of any one of claims 28-30, wherein each of the droplets comprises, on
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`average, less than five target nucleic acids.
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`67.|The methodof any one of claims 28-30, wherein the oil-immiscible fluid comprisesat
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`least one surfactant and glycerol.
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`68.
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`The method of any one of claims 28-30, wherein the oil-immiscible fluid comprisesat
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`least one surfactant, at least one antimicrobial agent and glycerol.
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`69.|The method of any one of claims 28-30, wherein the droplets flow through the output
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`flow path serially and substantially centered.
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`70.|The methodof any one of claims 28-30, wherein the droplets comprise a nucleic acid.
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`71.|The methodof any one of claims 28-30, wherein the droplets comprise a product of a
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`nucleic acid amplification reaction.
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`72.
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`The method of any one of claims 28-30, wherein the droplets are emulsified droplets.
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`73.|The method of any one of claims 28-30, wherein each of the droplets comprises, on
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`average, less than five target nucleic acids.
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