Droplet actuator devices, systems, and methods
First Claim
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1. A method of detecting a target nucleic acid in a sample, the method comprising:
- (a) providing a set of nucleic acid amplification reaction droplets, each droplet comprising a portion of the sample;
(b) treating two or more subsets of the amplification reaction droplets under conditions for amplifying the target nucleic acid to yield corresponding subsets of amplified droplets comprising amplified nucleic acid, wherein each subset of the amplification reaction droplets;
(i) comprises one or more of the amplification reaction droplets;
(ii) is treated under different conditions for amplifying the target nucleic acid;
(c) preparing the amplified droplets for detection;
(d) detecting a signal from the amplified droplets;
(e) based on the signal, determining the amount of the amplified nucleic acid present in the amplified droplets and/or the sample.
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Abstract
The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.
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Citations
79 Claims
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1. A method of detecting a target nucleic acid in a sample, the method comprising:
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(a) providing a set of nucleic acid amplification reaction droplets, each droplet comprising a portion of the sample; (b) treating two or more subsets of the amplification reaction droplets under conditions for amplifying the target nucleic acid to yield corresponding subsets of amplified droplets comprising amplified nucleic acid, wherein each subset of the amplification reaction droplets; (i) comprises one or more of the amplification reaction droplets; (ii) is treated under different conditions for amplifying the target nucleic acid; (c) preparing the amplified droplets for detection; (d) detecting a signal from the amplified droplets; (e) based on the signal, determining the amount of the amplified nucleic acid present in the amplified droplets and/or the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79)
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Specification