Legal claims defining the scope of protection, as filed with the USPTO.
1. A method for performing digital polymerase chain reaction (dPCR) in a dPCR biological analysis system including a thermal cycler, a processor, a detector, and a display, the method comprising: partitioning a biological sample volume including a plurality of target nucleic acids into a plurality of partitions, wherein at least one partition includes at least one target nucleic acid; amplifying, with the thermal cycler, the target nucleic acids in the plurality of partitions; generating, by the processor, a model for volume variation of the plurality of partitions based on a normal distribution of effective load volumes of the biological sample volume in the plurality of partitions; detecting, by the detector, the amplified target nucleic acids to determine a number of partitions including at least one target nucleic acid; calculating, by the processor, a concentration of target nucleic acids in the biological sample based on the model for volume variation and the number of partitions including at least one target nucleic acid; and displaying, on the display, the concentration of target nucleic acids in the biological sample.
2. The method of claim 1 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation: C = v 0 - v 0 2 + 2 σ 2 ln P ( neg ) σ 2 .
3. The method of claim 1 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation: P ( neg ) = erfc [ - 1 2 ( v 0 σ - C σ ) ] erfc [ - 1 2 ( v 0 σ ) ] e - Cv 0 + 1 2 σ 2 C 2 .
4. The method of claim 1 , further comprising: amplifying the target nucleic acids to determine the number of partitions including at least one target nucleic acid.
5. The method of claim 1 , wherein the model for volume variation is: P ( neg ) = erfc [ - 1 2 ( v 0 σ - C σ ) ] erfc [ - 1 2 ( v 0 σ ) ] e - Cv 0 + 1 2 σ 2 C 2 .
7. The method of claim 1 , wherein the plurality of partitions is a plurality of reaction sites.
8. The method of claim 1 , wherein the plurality of partitions is a plurality of throughholes.
9. The method of claim 1 , wherein the plurality of partitions is a plurality of droplets.
10. A system for performing digital polymerase chain reaction (dPCR), the system comprising: a device configured to partition a biological sample volume including a plurality of target nucleic acids into a plurality of partitions, wherein at least one partition includes at least one target nucleic acid; a thermal cycler to amplify the plurality of target nucleic acids; a detector for detecting the number of partitions including at least one target nucleic acid; a memory; and a processor configured to: generate a concentration of target nucleic acids in the biological sample based on a model for volume variation and the number of partitions including at least one target nucleic acid, wherein the model is based on a normal distribution of effective load volume of each of the biological sample volume in the plurality of partitions; and a display to display the concentration of target nucleic acids in the biological sample.
11. The system of claim 10 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation: C = v 0 - v 0 2 + 2 σ 2 ln P ( neg ) σ 2 .
12. The system of claim 10 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation: P ( neg ) = erfc [ - 1 2 ( v 0 σ - C σ ) ] erfc [ - 1 2 ( v 0 σ ) ] e - Cv 0 + 1 2 σ 2 C 2 .
13. The system of claim 10 , further comprising: an amplification apparatus configured to amplify the target nucleic acids to determine the number of partitions including at least one target nucleic acid.
14. The system of claim 10 , wherein the model for volume variation is: P ( neg ) = erfc [ - 1 2 ( v 0 σ - C σ ) ] erfc [ - 1 2 ( v 0 σ ) ] e - Cv 0 + 1 2 σ 2 C 2 .
16. The system of claim 10 , wherein the plurality of partitions is a plurality of reaction sites.
17. The system of claim 10 , wherein the plurality of partitions is a plurality of throughholes.
18. The system of claim 10 , wherein the plurality of partitions is a plurality of droplets.
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August 20, 2019
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