Silicone surfactants for emulsion assays

Patent metadata

Field	Value
Publication number	US-10150786-B2
Application number	US-201715697313-A
Country	US
Kind code	B2
Filing date	Sep 6, 2017
Priority date	Mar 15, 2013
Publication date	Dec 11, 2018
Grant date	Dec 11, 2018

Abstract

System, including methods and compositions, for making and using emulsions that include a silicone oil and a silicone surfactant. The emulsions may include aqueous droplets disposed in a continuous phase that includes a silicone oil and a silicone surfactant. The aqueous droplets may contain an analyte, optionally at partial occupancy, and/or a luminescent (e.g., photoluminescent) reporter. An assay of the analyte may be performed with the droplets. In some cases, signals may be detected from the droplets, and a characteristic of the analyte, such as an analyte level or activity, may be determined based on the signals.

First claim

We claim:

1. A method of performing an assay, the method comprising:
(A) forming an emulsion including aqueous droplets dispersed in a continuous phase, the aqueous droplets containing an analyte, the continuous phase including a silicone oil and a silicone surfactant, the silicone surfactant having the formula

the formula

or the formula

wherein
each g is independently 5-200;
each h is independently 1-5;
each i is independently 1-5;
each j is independently 15-50;
each k is independently 2-100; and
(B) detecting a signal related to the analyte from the aqueous droplets.

2. The method of claim 1 , wherein only a subset of the aqueous droplets contains the analyte.

3. The method of claim 1 , wherein the analyte is a nucleic acid.

4. The method of claim 1 , wherein the step of detecting a signal includes a step of detecting light.

5. The method of claim 4 , wherein the aqueous droplets contain a luminescent reporter, and wherein the step of detecting light includes a step of detecting light from the luminescent reporter.

6. The method of claim 5 , wherein the luminescent reporter includes a dye-labeled probe.

7. The method of claim 5 , wherein the luminescent reporter includes an intercalating dye.

8. The method of claim 1 , further comprising a step of performing an enzyme-catalyzed reaction in the aqueous droplets.

9. The method of claim 1 , wherein the silicone oil includes octamethyl trisiloxane, cyclopentasiloxane, or a trimethylsiloxysilicate at a concentration of about 0.1% to 5%.

10. The method of claim 1 , wherein the silicone oil includes one or more of octamethyl trisiloxane and polydimethylsiloxane.

11. The method of claim 1 , wherein the silicone surfactant is present in the continuous phase at a concentration of about 0.1% to 10% by weight.

Assignees

• Bio Rad Laboratories

Inventors

• Chia Erin R
• Hiddessen Amy L
• Hindson Benjamin J
• Lowe Adam
• Han Chunxiao

Classifications

• C08G77/50

  by carbon linkages · CPC title

• C07F7/184

  Chemistry & Metallurgy · mapped topic

• C08L83/12

  containing polyether sequences · CPC title

• C08L83/14

  in which at least two but not all the silicon atoms are connected by linkages other than oxygen atoms (C08L83/10 takes precedence) · CPC title

• C08G77/46 Primary

  containing polyether sequences · CPC title