Polymerase chain reaction method, polymerase chain reaction droplet device, and polymerase chain reaction droplet device array

• US 8,389,273 B2
• Filed: 02/10/2010
• Issued: 03/05/2013
• Est. Priority Date: 12/29/2009
• Status: Active Grant

• Current Assignee: National Applied Research Laboratories (Government of The Republic of China)
• Original Assignee: National Applied Research Laboratories (Government of The Republic of China)
• Inventors: Yu, Chih-Sheng, Tseng, Fan-Gang, Hu, Yi-Chiuem
• Primary Examiner(s): Woolwine, Samuel
• Assistant Examiner(s): Pande, Suchira
• Application Number:
  US12/703,209
• Publication Number:
  US 20110159547A1
• Time in Patent Office: 1,119 Days
• Field of Search: None
• US Class Current: 435/287.2
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• 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

Abstract

The present invention discloses a polymerase chain reaction (PCR) method, a PCR droplet device and a PCR droplet device array. The steps of the method comprise that a liquid comprising an analyzer is dropped on the heating coil disposed on the droplet device to form a droplet, then dropping a hydrophobic solution to prevent the droplet from evaporating. When an electric current or a voltage is supplied through at least one conducting wire to heat the heating coil, the inside of the droplet can generate buoyancy to drive the analyzer to move to the top of the inside of the droplet. Subsequently, the analyzer is moved to a periphery of the inside of the droplet so as to form a thermal cycle. Therefore the template is amplified by recycling the thermal cycle.

Claims

1. A polymerase chain reaction (PCR) droplet device, comprising:

a cooling plate;

a substrate disposed on the cooling plate;

a first containing area disposed on the substrate to contain a liquid comprising an analyzer;

a second containing area disposed on the substrate, and the second containing area surrounding the first containing area to contain a hydrophobic solution, wherein the hydrophobic solution is located at a surface of the liquid;

a heating coil disposed on a center of the first containing area and the heating coil is located inside the first containing area;

at least one conducting wire disposed on the substrate and connected with the heating coil; and

a plurality of sensors disposed around the heating coil to detect temperature changes of the heating coil;

wherein, the analyzer comprises a template, a primer, a deoxyribonucleotide triphosphate (dNTP) and a polymerase;

the at least one conducting wire is applied with an electric current or a voltage to heat the heating coil such that buoyancy in an inside of the liquid is generated, thereby driving the analyzer to move to a top of the inside of the liquid and subsequently to move to a periphery of the inside of the liquid so as to form a thermal cycle; and

the template is amplified by recycling the thermal cycle;

wherein, a metal electrical conductivity of the heating coil is higher than a metal electrical conductivity of the at least one conducting wire.

2. The PCR droplet device as claimed in claim 1, wherein the first containing area is formed by a first partition surrounding a periphery of the heating coil.

3. The PCR droplet device as claimed in claim 2, wherein a distance between the first partition and the heating coil is in a range of 0.5 to 2.5 mm.

4. The PCR droplet device as claimed in claim 3, wherein the second containing area is formed by a second partition surrounding a periphery of the first partition.

5. The PCR droplet device as claimed in claim 4, wherein a distance between the second partition and the heating coil is in a range of 1 to 3 mm.

6. The PCR droplet device as claimed in claim 1, wherein the first containing area is coated with a hydrophilic material.

7. The PCR droplet device as claimed in claim 6, wherein a functional group of the hydrophilic material comprises a hydrogen group, a carbonyl group, a carboxyl group, hydroxyl group, a sulfonic acid group, or an amino group.

8. The PCR droplet device as claimed in claim 7, wherein the second containing area is coated with a hydrophobic material.

9. The PCR droplet device as claimed in claim 8, wherein the hydrophobic material comprises epoxide, cyclic acetal, or styrene.

10. The PCR droplet device as claimed in claim 1, wherein the hydrophobic solution comprises mineral oil.

11. The PCR droplet device as claimed in claim 1, wherein a material of the substrate comprises silica, glass, nylon, polymer, or ceramic.

12. The PCR droplet device as claimed in claim 1, wherein a material of the heating coil comprises silver, copper, gold, platinum, aluminum, iron, stannum, lead or a combination thereof.

13. The PCR droplet device as claimed in claim 12, wherein a material of the at least one conducting wire comprises silver, copper, gold, platinum, aluminum, iron, stannum, lead or a combination thereof.

14. The PCR droplet device as claimed in claim 1, wherein the template comprises deoxyribonucleic acid (DNA) or ribonucleic acid (RNA).

15. The PCR droplet device as claimed in claim 1, wherein a shape of the heating coil comprises a circle, an angle, or an irregular shape.

16. The PCR droplet device as claimed in claim 1, wherein the liquid further comprises glycerol.

17. A polymerase chain reaction (PCR) droplet device array comprising a plurality of the PCR droplet device as claim 1, and the plurality of the PCR droplet devices disposed in an array mode.