Droplet-based nucleic acid amplification method and apparatus

• US 9,139,865 B2
• Filed: 11/19/2012
• Issued: 09/22/2015
• Est. Priority Date: 04/18/2006
• Status: Active Grant

• Current Assignee: Duke University, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Duke University, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Pollack, Michael G., Paik, Philip, Pamula, Vamsee K.
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US13/680,979
• Publication Number:
  US 20130092539A1
• Time in Patent Office: 1,037 Days
• Field of Search: 422502-504, 422/509, 422/518, 204600-609, 700/266, 700/283, 700/286
• US Class Current: 1/1
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1894 Cooling means; Cryo cooling
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• 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 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: C12Q 2565/301 Pyrophosphate (PPi)
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

Aspects of embodiments of the invention relate to a simulator including a visual display capable of outputting to a user a display one or more effects of a command series selected and a system including a droplet microactuator electronically coupled to and controlled by a processor capable of executing instructions, the droplet microactuator including a substrate comprising electrodes for conducting droplet operations. Further aspects of embodiments of the invention relate to a droplet operations troubleshooting apparatus. Other aspects of embodiments of the invention relate to a computer implemented method of displaying simulated microactuator droplets.

Claims

1. A system for simulating droplet operations on a droplet microactuator comprising:

(a) an output device configured to output to a user a display of one or more effects of a selected command series;

(b) a system comprising a droplet microactuator and a processor capable of executing instructions, wherein the droplet microactuator is electronically coupled to and controlled by the processor, the droplet microactuator comprises a substrate comprising electrodes for conducting electrowetting-mediated droplet operations, and wherein the system is configured and programmed to display a simulated execution of a subroutine on a simulated droplet on the droplet microactuator, wherein the simulated execution of the subroutine on the simulated droplet is displayed via the output device to the user; and

wherein the simulated execution comprises a map simulating the position of each of the electrodes and differentiates the electrodes in their simulated activated and unactivated states on the map by appearance.

2. The system for simulating of claim 1, wherein the output device comprises a screen.

3. The system for simulating of claim 1, further comprising executable instructions that are stored on a computer readable medium.

4. The system for simulating of claim 1, further comprising software, wherein the software executes steps of a subroutine but does not send an electrical signal to the droplet microactuator.

5. The system for simulating of claim 1, wherein the droplet microactuator further comprises one or more temperature control means arranged for heating a region of the droplet microactuator such that a droplet can be transported into the region for heating.

6. The system for simulating of claim 1, further comprising a reservoir.

7. The system for simulating of claim 1, wherein the substrate comprises an electrowetting substrate.

8. The system for simulating of claim 2, wherein the processor comprises executable code programmed to display on the screen an actual effect of a program resulting from the one or more effects of a command series selected.

9. The system for simulating of claim 2, wherein the processor comprises executable code programmed to display on the screen a simulated effect of a droplet microactuator resulting from the one or more effects of a command series selected.

10. The system for simulating of claim 2, wherein the processor comprises executable code programmed to display on the screen a simulated execution of a subroutine on a droplet microactuator map resulting from the one or more effects of a command series selected.

11. An apparatus for droplet operations simulations comprising:

(a) a system comprising a droplet microactuator and a processor capable of executing instructions, wherein the droplet microactuator is electronically coupled to and controlled by the processor, and wherein the droplet microactuator comprises a substrate comprising electrodes for conducting droplet operations and one or more temperature control means arranged for heating a region of the droplet microactuator such that a droplet can be transported via the electrodes into the region for heating, and wherein the system is configured and programmed to display a simulated execution of a subroutine on a simulated droplet on the droplet microactuator;

wherein the simulated subroutine on the simulated droplet is displayed via an output device to a user; and

(b) the output device is configured to display to the user effects of a selected command series.

12. The apparatus of claim 11, wherein the substrate comprises an electrowetting substrate.

13. The apparatus of claim 11, further including a filler fluid.

14. The apparatus of claim 11, further including a filler fluid comprising oil.

15. The apparatus of claim 11, further including a detector.