Patent ID: 11884973
Assignee: HITACHI, LTD.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 5:
6. The method of claim 5, wherein the plurality of fluorescent labeled probes further comprises a third fluorescent labeled probe comprises a second fluorescent dye associated with a second fluorescence color and the fluorescence intensity associated with the plurality of fluorescent labeled probes further comprises a second fluorescence intensity associated with the second fluorescence color, wherein the third fluorescent labeled probe is configured for hybridization to a wild type target DNA, wherein the first reference Tm and the second reference Tm are different from a third reference Tm calculated for the third fluorescent labeled probe and the wild type target DNA of the plurality of target DNAs in bulk, and wherein:
the fourth step further comprises determining,
that the wild type target DNA is not contained in droplets in a fifth set of droplets in the plurality of droplets based on (1) the measured second fluorescence intensity associated with the second fluorescence color being below a second threshold fluorescence intensity associated with the second fluorescence color or (2) the measured second fluorescence intensity being above the second threshold fluorescence intensity and a third calculated Tm of the at least one calculated Tm being out of a third predetermined range of Tm around the third reference Tm;

the fifth step further comprises counting:
a fourth number of droplets in a sixth set of droplets in the plurality of droplets, wherein the sixth set of droplets are determined to contain the wild type target DNA based on (1) the measured second fluorescence intensity being equal to, or larger than, the second threshold fluorescence intensity and (2) the third calculated Tm being in the third predetermined range of Tm around the third reference Tm; and

the sixth step further comprises displaying, on the monitor, the fourth number of droplets in the sixth set of droplets that contain the wild type target DNA, wherein the third number of droplets is a third number of droplets in the intersection of the first set of droplets, the second set of droplets, and the fifth set of droplets that do not contain at least one of the first target DNA, the second target DNA, or the wild type target DNA.