Devices for fast DNA replication by polymerase chain reactions (PCR)

• US 6,428,987 B2
• Filed: 12/12/2000
• Issued: 08/06/2002
• Est. Priority Date: 04/23/1997
• Status: Expired due to Fees

• Current Assignee: Bruker Daltonik GmbH (Bruker Corporation)
• Original Assignee: Bruker Daltonik GmbH (Bruker Corporation)
• Inventors: Franzen, Jochen
• Primary Examiner(s): Benzion, Gary
• Assistant Examiner(s): Tung, Joyce
• Application Number:
  US09/735,372
• Publication Number:
  US 20010000752A1
• Time in Patent Office: 602 Days
• Field of Search: 435/912, 435/290, 435/316, 422/196, 422/197, 422/68.1, 422/57, 422/81.12
• US Class Current: 435/91.2
• 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 instruments for fast, selective replication of deoxyribonucleic acid (DNA) from biomaterial through 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 microsystem technology.

Claims

1. Apparatus for use in replicating DNA located in a polymerase chain reaction (PCR) fluid, the apparatus comprising:

a housing having a flow path through which the reaction fluid may flow, the flow path including a set of reaction chambers comprising a plurality of capillaries into which fluid flowing through the flow path is divided to flow simultaneously before being recombined, such that a flow rate of the reaction solution through the parallel capillaries is lower than elsewhere in the flow path; and

heating apparatus that provides heat to the flow path such that at least three different temperatures affect the reaction solution, the solution being subjected to at least one of the three temperatures while simultaneously present in the capillaries.

2. Apparatus according to claim 1, wherein the capillaries each have a diameter below 400 micrometers.

3. Apparatus according to claim 1, wherein the volume of each reaction chamber is less than one microliter.

4. Apparatus according to claim 1, wherein the capillary reaction chambers are each microfabricated from silicon, and the surface of the capillaries is coated with a coating material that does not inhibit PCR.

5. Apparatus according to claim 4, wherein the coating material has been bonded chemically to a surface of the silicon.

6. Apparatus according to claim 4, wherein the coating material comprises a chromatographic coating.

7. Apparatus according to claim 4, wherein the coating material comprises nonpolar phase Carbowax.

8. Apparatus according to claim 4, wherein the coating material comprises glycoproteins, proteins or lipoproteins.

9. Apparatus according to claim 1, wherein the capillaries comprise polymer plastics.

10. Apparatus according to claim 1, wherein the capillaries comprise polyethylene or polypropylene.

11. Apparatus according to claim 1, wherein capillaries comprise, at least in part, a material with a high thermal conductivity.

12. Apparatus according to claim 1 wherein the three temperatures correspond to temperatures appropriate for the phases of PCR melting, primer attachment and reconstruction.

13. Apparatus according to claim 12 wherein a reaction solution passing through the flow path is at the reconstruction temperature while in the capillaries.

14. Apparatus according to claim 1 wherein the heating apparatus comprises fluid cooling means that can be brought into thermal contact with the capillaries to change the temperature of the reaction solution in the capillaries.

15. Apparatus according to claim 1 wherein the flow rate through the capillaries is no greater than a value equal to ten times the diameter of each capillary per second.

16. Apparatus according to claim 1 wherein the plurality of capillaries is a first set of capillaries and wherein the housing comprises multiple sets of capillaries through which the reaction fluid must flow.

17. Apparatus according to claim 1 further comprising a flow inducer for inducing flow of the reaction fluid through the flow path.

18. Apparatus according to claim 17 wherein the flow inducer provides pressure pulses to the fluid in the flow path.

19. Apparatus for use in performing the steps of PCR melting, primer attachment and reconstruction for replicating DNA located in a polymerase chain reaction (PCR) fluid, the apparatus comprising:

a housing having a flow path through which the reaction fluid may flow, the flow path including a set of reaction chambers comprising a plurality of capillaries into which fluid flowing through the flow path is divided to flow simultaneously before being recombined, such that a flow rate of the reaction solution through the parallel capillaries is lower than elsewhere in the flow path;

heating elements adjacent to the fluid path to change the reaction solution temperature from the primer attachment temperature to the temperatures for PCR reconstruction and melting; and

gaseous, liquid or solid cooling means that can be brought into thermal contact with the flow path to reduce the temperature of the reaction solution in the flow path.

20. Apparatus according to claim 19, wherein the flow path is formed, at least in part, by a surface that is thinner than one millimeter.

21. Apparatus according to claim 19, wherein a portion of the flow path comprises a single, convoluted capillary.

22. Apparatus according to claim 19 wherein the plurality of capillaries is a first set of capillaries and wherein the housing comprises multiple sets of capillaries through which the reaction fluid must flow.