Method and device for conducting biochemical or chemical reactions at multiple temperatures

• US 9,452,433 B2
• Filed: 05/29/2014
• Issued: 09/27/2016
• Est. Priority Date: 05/11/2005
• Status: Expired due to Fees

• Current Assignee: Duke University, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Duke University, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Shenderov, Alexander, Pollack, Michael G.
• Primary Examiner(s): PRIEST, AARON A
• Application Number:
  US14/290,057
• Publication Number:
  US 20140329307A1
• Time in Patent Office: 852 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/089 Virtual walls for guiding liquids
• CPC Class Codes: B01L 2300/1816 using induction heating
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1872 Infrared light
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones

Abstract

Methods and devices for conducting chemical or biochemical reactions that require multiple reaction temperatures are described. The methods involve moving one or more reaction droplets or reaction volumes through various reaction zones having different temperatures on a microfluidics apparatus. The devices comprise a microfluidics apparatus comprising appropriate actuators capable of moving reaction droplets or reaction volumes through the various reaction zones.

Claims

1. A device for PCR amplifying nucleic acids, the device comprising:

(a) a first substrate and a second substrate separated to form a gap, an electrowetting path or array comprising a plurality of electrowetting electrodes on one or both sides of the gap and two or more reaction zones having different temperatures;

(b) a droplet on the electrowetting path or array comprising a nucleic acid and amplification reagents, wherein the droplet is disposed between the first and second substrates and in contact with both the first and second substrates; and

(c) a filler fluid surrounding the droplet and filling the gap.

2. The device of claim 1 wherein the plurality of electrowetting electrodes is configured to move the droplet through each of the two or more reaction zones.

3. The device of claim 1 wherein the reaction zones comprise reaction zones having temperatures suitable for amplifying the nucleic acid.

4. The device of claim 3 wherein the reaction zones comprise reaction zones having temperatures selected to effect denaturing of the nucleic acids, annealing of primers to the nucleic acids, and/or polymerization of the nucleic acids.

5. The device of claim 1 wherein at least one of the two or more reaction zones is configured to be kept at a constant temperature.

6. The device of claim 5 wherein the at least one of the two or more reaction zones is configured to be kept at a constant temperature by a resistive heating mechanism.

7. The device of claim 5 wherein the at least one of the two or more reaction zones is configured to be kept at a constant temperature by an inductive heating mechanism.

8. The device of claim 5 wherein the at least one of the two or more reaction zones is configured to be kept at a constant temperature by an infrared heating mechanism.

9. The device of claim 1, wherein the filler fluid comprises silicone oil.

10. The device of claim 1, comprising a multitude of parallel electrowetting paths.

11. The device of claim 1, wherein the electrowetting path or array comprises a detection zone.

12. The device of claim 1, wherein the electrowetting electrodes are on both sides of the gap.