Multiplexed digital assays with combinatorial use of signals
DCFirst Claim
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1. A method of performing a multiplexed digital amplification assay, the method comprising:
- amplifying more than R targets in partitions;
creating R signals representative of light detected in R different wavelength regimes from the partitions, where R≧
2; and
calculating an average level of each target in the partitions based on the R signals, wherein the level calculated accounts for a coincidence of all possible combinations of the more than R targets in the same individual partitions;
wherein the more than R targets include three targets, and wherein the average levels of the three targets are calculated based on light detected from only two fluorophores associated with probes that bind to amplicons of the three targets during amplification.
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Abstract
System, including methods, apparatus, and compositions, for performing a multiplexed digital assay on a greater number of targets through combinatorial use of signals.
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Citations
15 Claims
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1. A method of performing a multiplexed digital amplification assay, the method comprising:
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amplifying more than R targets in partitions; creating R signals representative of light detected in R different wavelength regimes from the partitions, where R≧
2; andcalculating an average level of each target in the partitions based on the R signals, wherein the level calculated accounts for a coincidence of all possible combinations of the more than R targets in the same individual partitions; wherein the more than R targets include three targets, and wherein the average levels of the three targets are calculated based on light detected from only two fluorophores associated with probes that bind to amplicons of the three targets during amplification. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method of performing a multiplexed digital amplification assay, the method comprising:
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amplifying targets in partitions; detecting light from the partitions; and calculating levels of at least three of the targets based on light detected from only two fluorophores, wherein the levels calculated account for a coincidence of all possible combinations of the at least three targets in the same individual partitions. - View Dependent Claims (13, 14, 15)
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Specification