Method and devices for extremely fast DNA replication by polymerase chain reactions (PCR)

• US 6,180,372 B1
• Filed: 03/27/1998
• Issued: 01/30/2001
• Est. Priority Date: 04/23/1997
• Status: Expired due to Term

• Current Assignee: Bruker-Franzen Analytik GmbH
• Original Assignee: Bruker Daltonik GmbH (Bruker Corporation)
• Inventors: Franzen, Jochen
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): Tung, Joyce
• Application Number:
  US09/049,646
• Time in Patent Office: 1,040 Days
• Field of Search: 435/91.2, 435/290, 435/316, 435/91.1, 422/196, 422/197, 422/68.1, 422/57, 422/81.12
• US Class Current: 435/91.1
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• 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: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2527/113 Time
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: Y10S 977/775 Nanosized powder or flake, e.g. nanosized catalyst
• CPC Class Codes: Y10S 977/783 Organic host/matrix, e.g. lipid
• CPC Class Codes: Y10S 977/788 Of specified organic or carbon-based composition
• CPC Class Codes: Y10S 977/80 Nucleic acid, e.g. DNA or RNA
• CPC Class Codes: Y10S 977/924 : using nanostructure as support of dna analysis

Abstract

The invention concerns methods and instruments for fast, selective replication of deoxyribonucleic acid (DNA) from biomaterial through the known polymerase chain reaction (PCR), working in individual duplication thermocycles. The invention consists of extremely brief cycle times of only a few seconds for the PCR reactions, generated, on the one hand, by reaction chambers for the reception of the reaction solution constructed of a pattern of fine capillaries in close proximity to heating and cooling elements in order to optimally accelerate the temperature setting in the reaction solution for the three temperature phases of the PCR duplication cycles and, on the other hand, by keeping the flow rates in the capillaries to a minimum during the amplification phase so that the polymerase reaction is not disturbed. The capillary pattern can be simply produced by means of microsystern technology.

Claims

1. A method of replicating DNA located in a polymerase chain reaction (PCR) solution, the method comprising:

transferring the reaction solution to a set of reaction chambers comprising a plurality of fine capillaries that may be filled simultaneously; and

moving the reaction solution through three different temperature zones for PCR melting, primer attachment and reconstruction, respectively, wherein the solution is subjected to at least one of the three temperatures while simultaneously present in the plurality of capillaries, a local flow rate of the reaction solution through the capillaries being lower than elsewhere in a flow path of the reaction solution.

2. A method according to claim 1 wherein the reaction solution is at the reconstruction temperature while in the capillaries.

3. A method according to claim 1, wherein the capillaries have diameters below 400 micrometers.

4. A method according to claim 2, wherein the local flow rate through the capillaries is no greater than a value equal to ten times the diameter of each capillary per second.

5. A method according to claim 1, wherein the volume of each capillary is less than one microliter.

6. A method according to claim 1, wherein the capillary reaction chambers are microfabricated from silicon, and wherein the method further comprises coating the surface of the capillaries with a coating material that and does not inhibit PCR.

7. A method according to claim 6 wherein coating the surface of the capillaries further comprises chemically bonding molecules of the coating to the surface of the silicon.

8. A method according to claim 7 wherein chromatographic phases are used for the coating.

9. A method according to claim 7, wherein the coating comprises polyethylene glycol.

10. A method according to claim 7, wherein glycoproteins of cell membranes, proteins or lipoproteins are used for coating the capillary surfaces.

11. A method according to claim 1 further comprising microfabricating the capillary reaction chambers from polymer plastics.

12. A method according to claim 1 further comprising microfabricating the capillary reaction chambers from polyethylene or polypropylene.

13. A method according to claim 11, wherein the polymer plastics are filled with particles from a thermally highly conductive metal.

14. A method according to claim 13 wherein said particles comprise silver powder.

15. A method according to claim 11 further comprising coating inner surfaces of the capillaries to deactivate the surfaces.

16. A method according to claim 1 wherein the solution is subjected to each of the three temperatures while present in the plurality of capillaries.

17. A method according to claim 1, wherein the capillaries are located in a thin, microfabricated membrane.

18. A method according to claim 1, wherein the reaction chamber comprises a zig-zag capillary.

19. A method according to claim 1, wherein each of the capillaries has a substantially equal flow resistance.

20. A method according to claim 1 wherein moving the reaction fluid comprises moving the reaction fluid by applying pressure pulses to it.

21. A method according to claim 17 wherein a thickness of the membrane is less than one millimeter.

22. A method according to claim 17 further comprising providing heat to solution in the capillaries with heating elements that contact a surface of the membrane.

23. A method according to claim 17 further comprising providing cooling to solution in the capillaries with cooling elements that contact a surface of the membrane.