IP Library › Granted Patent US 12,241,116
Granted Patent B2
US 12,241,116 · App. 18/536,935 · Granted Mar 4, 2025

Digital analyte analysis

Inventors: Jonathan William Larson (Chelsea, MA); Qun Zhong (Lexington, MA); Darren Roy Link (Lexington, MA)
Assignee: BIO-RAD LABORATORIES, INC.
C12Q1/6816C12Q1/6827C12Q1/686C12Q1/6883C12Q1/6886C12Q2600/172
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Quick Facts
Patent No.
US 12,241,116
App. No.
18/536,935
Granted
Mar 4, 2025
Kind
B2
Abstract

The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, methods of the invention involve forming sample droplets containing, on average, a single target nucleic acid, amplifying the target in the droplets, excluding droplets containing amplicon from the target and amplicon from a variant of the target, and analyzing target amplicons.

Claims (17)

1. A composition for use in a digital PCR assay, the composition comprising:

a plurality of primers and a plurality of probes,

wherein a first probe of the plurality of probes is configured to bind to a first nucleic acid target,

wherein the first probe of the plurality of probes comprises a first label,

wherein a second probe of the plurality of probes is configured to bind to a second nucleic acid target,

wherein the second probe of the plurality of probes comprises a second label,

wherein the first label is a first fluorophore and the second label is a second fluorophore, wherein the first fluorophore and the second fluorophore are the same color, wherein the composition includes the first probe at a first concentration and the second probe at a second concentration different from the first concentration, and

wherein the first fluorophore and the second fluorophore are detectable in a detection channel.

2. The composition of claim 1 , wherein when the composition is used in the digital PCR assay, the first label is detectable by a first intensity of the color in the detection channel and the second label is detectable by a second intensity, different from the first intensity, of the color in the detection channel.

3. The composition of claim 2 , wherein the difference in intensity between the first intensity and the second intensity is based on the first concentration of the first probe and the second concentration of the second probe.

4. The composition of claim 3 , wherein the difference in intensity between the first intensity and the second intensity is further based on a first concentration of a first primer that binds to the first nucleic acid target and a second concentration of a second primer that binds to the second nucleic acid target.

5. The composition of claim 3 , wherein the difference in intensity between the first intensity and the second intensity is further based on a first amplification efficiency of the first nucleic acid target and a second amplification efficiency of the second nucleic acid target.

6. The composition of claim 3 , wherein the difference in intensity between the first intensity and the second intensity is further based on use of one or more blocking oligomer nucleotides.

7. The composition of claim 1 , wherein the composition comprises a third probe of the plurality of probes, wherein the third probe is configured to bind to the first nucleic acid target, and wherein the third probe comprises a third label that is different from the first label.

8. The composition of claim 1 , wherein the plurality of primers comprises at least a first primer that binds to the first nucleic acid target and at least a second primer that binds to the second nucleic acid target.

9. The composition of claim 8 , wherein the first primer is present in the composition at a first concentration, and the second primer is present in the composition at a second concentration different from the first concentration.

10. The composition of claim 2 , wherein the first intensity is a first fluorescence intensity and the second intensity is a second fluorescence intensity.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 12, 2024
From: LARSON, JONATHAN WILLIAM; ZHONG, QUN; LINK, DARREN ROY
To: RAINDANCE TECHNOLOGIES, INC.
Reel/Frame 066439/0075 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 12, 2024
From: RAINDANCE TECHNOLOGIES, INC.
To: BIO-RAD LABORATORIES, INC.
Reel/Frame 066439/0246 →
Continuity (10)
Continuation 17574780 · Jan 13, 2022
Continuation 15688250 · Aug 28, 2017
Continuation 15019355 · Feb 9, 2016
Continuation 13773868 · Feb 22, 2013
Continuation 13026120 · Feb 11, 2011
Provisional Application 61388937 · Oct 1, 2010
Provisional Application 61347158 · May 21, 2010
Provisional Application 61331490 · May 5, 2010
Provisional Application 61304163 · Feb 12, 2010
Related Publication 20240132939A1 · Apr 25, 2024
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