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US 2016/0108464 A1Published application
Multiplexed digital assays with combinatorial use of signals
AssigneeBio-Rad Laboratories, Inc.
InventorsSerge Saxonov, Simant Dube, Benjamin J. Hindson, Adam M. McCoy
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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.
Claims20
A method of determining an amount of target linkage, the method comprising:
- partitioning an aqueous phase containing a sample into a plurality of partitions, the sample including nucleic acid providing a first target and a second target;
- amplifying the first target and the second target in partitions;
- collecting amplification data from partitions;
- determining a number of partitions containing the first target and not the second target, a number of partitions containing the second target and not the first target, a number of partitions containing both targets, and a number of partitions containing neither target;
- and
- calculating a value indicating a percent linkage of the first target and the second target to one another in the sample using a plurality of the numbers.
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A composition for performing a multiplexed digital amplification assay, comprising:
- an emulsion including droplets, each droplet including a portion of a same aqueous phase;
- wherein the aqueous phase includes three probes and is configured to amplify a first target, a second target, and a third target,
- wherein each of the three probes is configured to detect amplification of a different one of the targets,
- wherein the three probes are labeled with a total of no more than two different fluorophores, and
- wherein each droplet of a subset of the droplets contains the first target and the second target but not the third target, each droplet of another subset of the droplets contains the first target and the third target but not the second target, each droplet of yet another subset of the droplets contains the second target and the third target but not the first target, and each droplet of still another subset of the droplets contains none of the targets.
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Dependencies are inferred from the claim text.