Polymerase chain reaction container and process for producing the same

• US 20040175713A1
• Filed: 01/30/2004
• Published: 09/09/2004
• Est. Priority Date: 07/05/2002
• Status: Active Grant

• Current Assignee: Panasonic Corporation (Panasonic Holdings Corporation)
• Original Assignee: Panasonic Corporation (Panasonic Holdings Corporation)
• Inventors: Nakatani, Masaya, Yukimasa, Tetsuo
• Granted Patent: US 7,384,782 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• 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: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

The present invention provides a kit employed for polymerase chain reaction. The kit has a cavity and a flow channel on a substrate. The flow channel is separated from the cavity by at least a barrier formed along the cavity. Such a structure allows the cavity to be filled with a sample solution even in minute quantities. At the same time, the structure can provide a sample solution with a rapid temperature-control. The structure can therefore contribute to accelerated polymerase chain reaction.

Claims

1. A polymerase chain reaction kit comprising:

a substrate on which a cavity, a flow channel for circulating a heating medium, and a barrier for separating the cavity and the flow channel are formed; and

at least one cover plate of a plurality of cover plates for sealing at least one of the cavity and the flow channel.

2. The polymerase chain reaction kit of claim 1, wherein the substrate has the flow channel and the cavity on a same surface.

3. The polymerase chain reaction kit of claim 1, wherein the cavity is formed in a first surface of the substrate, whereas the flow channel is formed in a second surface opposite to the first surface of the substrate, the plurality of cover plates include a first cover plate for sealing the cavity, and a second cover plate for sealing the flow channel.

4. The polymerase chain reaction kit of claim 1, wherein the flow channel is formed on the substrate so as to run along at least a section of a side of the cavity.

5. The polymerase chain reaction kit of claim 3, wherein the flow channel is formed on the substrate so as to run along at least a section of a bottom of the cavity.

6. The polymerase chain reaction kit of claim 1, wherein at least a portion, which separates the cavity from the flow channel, of the barrier is formed of same material as that forming the substrate.

7. The polymerase chain reaction kit of claim 3, wherein at least a portion, which separates the cavity from the flow channel, of the barrier is formed of material having an etching rate smaller than that forming the substrate in an identical etching method.

8. The polymerase chain reaction kit of claim 1, wherein the substrate is made of silicon.

9. The polymerase chain reaction kit of claim 1, wherein the substrate is formed of any one of i) silicon, ii) silicon oxide, and iii) glass including silicon oxide, similarly, the cover plate is formed of any one of i) silicon, ii) silicon oxide, and iii) glass including silicon oxide.

10. The polymerase chain reaction kit of claim 1, wherein the substrate has a multi-layered structure formed of silicon layer and any one layer of i) silicon dioxide and ii) a glass substrate including silicon dioxide.

11. The polymerase chain reaction kit of claim 3, wherein material of at least a portion, which separates the cavity from the flow channel, of the barrier includes any one of i) silicon dioxide, ii) silicon nitride, iii) gold, iv) platinum, v) chrome, and vi) nickel.

12. The polymerase chain reaction kit of claim 1, wherein the cover plate contains any one of i) a sample-injection inlet connected to the cavity, ii) a flow-in hole connected to the flow channel, and iii) a flow-out hole connected to the flow channel.

13. The polymerase chain reaction kit of claim 3, wherein the first cover plate contains a sample-injection inlet connected to the cavity, while the second cover plate contains a flow-in hole and a flow-out hole connected to the flow channel.

14. The polymerase chain reaction kit of claim 12, wherein at least any one of i) the sample-injection inlet, ii) the flow-in hole, and iii) the flow-out hole, all of which are disposed in the cover plate, is formed into conic so as to become narrower on a side attached to the substrate.

15. The polymerase chain reaction kit of claim 1, wherein at least any one of i) a sample-injection inlet connected to the cavity, ii) a flow-in hole connected to the flow channel, and iii) a flow-out hole connected to the flow channel is disposed on a side section formed by bonding the substrate and the cover plate.

16. The polymerase chain reaction kit of claim 1, wherein the substrate and the cover plate are directly attached.

17. The polymerase chain reaction kit of claim 1, wherein the barrier has a thickness at most 100 μ

m.

18. The polymerase chain reaction kit of claim 1, wherein the cavity formed on the substrate is shaped into any one of oval and rectangular in a horizontal plane.

19. The polymerase chain reaction kit of claim 4, wherein the cavity formed on the substrate is shaped into any one of i) a meander structure with at least two bends, and ii) a spiral structure.

20. The polymerase chain reaction kit of claim 5, wherein the substrate contains the flow channel having a bottom area wider than that of the cavity.

21. A method of manufacturing a polymerase chain reaction kit comprising the steps of:

a) forming a cavity and a flow channel for circulating a heating medium separated from the cavity on a surface of a substrate;

b) attaching the substrate to at least one cover plate of a plurality of cover plates;

c) connecting the cavity to a sample-injection inlet exposed to outside; and

d) connecting the flow channel to a flow-in hole and a flow-out hole exposed to outside.

22. The method of manufacturing a polymerase chain reaction kit of claim 21, wherein the a) step employs at least a promotive gas and a suppressive gas for etching.

23. The method of manufacturing a polymerase chain reaction kit of claim 21, wherein the cavity and the flow channel are formed in a same surface of the substrate in the a) step.

24. The method of manufacturing a polymerase chain reaction kit of claim 21, wherein at least any one of the sample-injection inlet, the flow-in hole and the flow-out hole is formed in the cover plate.

25. The method of manufacturing a polymerase chain reaction kit of claim 21, wherein the a) step further includes the sub steps of:

a-1) forming the cavity on a first surface of the substrate; and

a-2) forming the flow channel on a second surface opposite to the first surface of the substrate,

and the plurality of cover plates include a first cover plate for sealing the cavity and a second cover plate for sealing the flow channel.

26. The method of manufacturing a polymerase chain reaction kit of claim 25, wherein the sample-injection inlet is formed in the first cover plate, while the flow-in hole and the flow-out hole are formed in the second plate.

27. The method of manufacturing a polymerase chain reaction kit of claim 25 further includes a step of e) forming a layer, which is formed of a material having an etching rate smaller than that of a material forming the substrate, on a inner wall of the cavity.

28. The method of manufacturing a polymerase chain reaction kit of claim 27, wherein the substrate is made of silicon, the layer is made of silicon dioxide, and the layer is formed by any one of i) thermal oxidization, ii) chemical vapor deposition (CVD), and iii) sputtering.

29. The method of manufacturing a polymerase chain reaction kit of claim 27, wherein the substrate is made of silicon, the layer is made of any one of i) gold, ii) platinum, iii) chrome, iv) nickel, and the layer is formed by any one of i) CVD, ii) sputtering, iii) vacuum evaporation, and iv) plating.

30. The method of manufacturing a polymerase chain reaction kit of claim 21, wherein the b) step employs any one of direct attachment and anodic attachment.

31. The method of manufacturing a polymerase chain reaction kit of claim 24, wherein any one of the sample-injection inlet, the flow-in hole and the flow-out hole is formed, by sand blasting, in at least one of the c) step and d) step.