Method and device for preparing samples for detecting a nucleotide sequence

• US 6,383,802 B1
• Filed: 12/11/2000
• Issued: 05/07/2002
• Est. Priority Date: 06/12/1998
• Status: Expired due to Fees

• Current Assignee: November Aktiengesellschaft Gesellschaft Fur Molekulare Medizin
• Original Assignee: November Aktiengesellschaft Gesellschaft Fur Molekulare Medizin
• Inventors: Bertling, Wolf
• Primary Examiner(s): Sisson, B. L.
• Application Number:
  US09/719,376
• Time in Patent Office: 512 Days
• Field of Search: 435/287.2, 435/6, 435/91.1, 435/91.2, 435/183, 536/23.1, 536/24.3, 536/24.33
• US Class Current: 435/287.2
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2527/101 Temperature
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels

Abstract

The invention relates to a method for preparing samples for detecting a nucleotide sequence by polymerase chain reaction (PCR), according to which a) an analysis solution is filled into at least one cavity (2) provided for in a support (1); b) a lid (3) configured complementary to the shape of the cavity (2) is placed onto the support (1) in such a way that the analysis solution is pushed at least partly into a gap (S) formed between the cavity (2) and the lid (3); and c) the gap (S) is sealed by means of at least one seal (5, 12) provided for near an opening in the cavity (2).

Claims

1. A device for preparing one or more samples for detecting a nucleotide sequence by means of polymerase chain reaction (PCR) where each sample is contacted with an analysis solution, said device comprising a support (1) with at least one cavity (2), a lid (3) which is configured complementary to the cavity (2) and which can be placed on the support (1) such that at least some of the analysis solution taken up by the cavity (2) can be displaced into a gap (2) formed between the cavity (2) and the lid (3) and with a seal (5, 12), arranged adjacent an opening of the cavity (2) for sealing the gap (G), wherein the cavity (2) has a surrounding lateral wall (LW) which widens conically toward the opening and the gap (G) has a width (W) of not more than 1 mm, an electrode (6) being integrally molded into the lid (3) and a primer being bound to an internal face (I) of the lid (3), which faces the cavity (2).

2. The device according to claim 1, further comprising a facility for cyclically heating and cooling the analysis solution.

3. The device according to claim 1, further comprising a facility for examining the fluorescence properties of at least one of the analysis solution and one of the primers.

4. The device according to claim 1, wherein the support is made of a translucent material.

5. The device according to claim 1, wherein the cavity has planar sections and has a flat bottom (B).

6. The device according to claim 5, further comprising means for exciting fluorescence between the bottom (B) and the internal face (I) of the lid (3).

7. The device according to claim 6, wherein radiation originating from the excitation means can be focused toward the internal face (I) of the lid (3).

8. The device according to claim 6, wherein the means (7) for exciting fluorescence is generated by a laser diode.

9. The device according to claim 6, further comprising a facility (8) for detecting the fluorescence.

10. The device according to claim 9, further comprising a facility for evaluating the detected fluorescence.

11. The device according to claim 9, further comprising a facility for shifting the support (1) relative to at least one of the means (7) for exciting the fluorescence and the detection facility (8).

12. The device according to claim 11, further comprising a facet-eye-like means for separately exciting and/or detecting the fluorescence between each bottom (B) and the internal face (I) of the corresponding lid (3).

13. The device according to claim 1, comprising a further seal (12), said further seal being provided on the support (1) and/or on the internal face (I) of the lid (3) in sections located between the cavities (2) or on projections (4) provided on the internal face (I) of the lid (3).

14. The device according to claim 1, wherein the support has 96 cavities (2) and the lid (3) 96 projections (4), said projections being complementary to the shape of the cavities (2).

15. The device according to claim 1, wherein the lid (3) is made of an electroconductive plastic material.

16. The device according to claim 15, wherein the plastic material is selected from the group consisting of polycarbonate, trimethylthiophene, triaminobenzene and polycarbene and at least sections of the internal face (I) of the lid (3) are provided with a substance which binds biomolecules.

17. The device according to claim 11, wherein the support (1) has a counterelectrode that is operatively linked with the lid electrode (6) so that an electrical field can be applied by means of which nucleotide sequence present in the analysis solution can be shifted to the internal face (I) and accumulated by field-inversion cycles.

18. The device according to claim 17, wherein a portion of said electrode (6) is exposed at said face (I) of said lid (3).

19. The device according to claim 11, wherein the analysis solution has a first primer added to it.

20. The device according to claim 19, wherein the analysis solution has a second primer added to it.

21. The device according to claim 11, wherein said primer is bound to the internal face (I) of the lid (3) facing the cavity (2), with its 5′

-terminal end.

22. The device according to claim 11, wherein at least sections of the lid (3) and/or the support (1) are black so that heat radiated at them is absorbed in an efficient manner.

23. The device according to claim 22, further comprising a facility for cyclically heating and cooling the analysis solution.

24. The device according to claim 23, further comprising a heating means (9), said heating means being selected from the group consisting of infra-red radiation heaters, resistance heating and means for passing a gas or a fluid around the cavity (2).

25. The device according to claim 23, further comprising means (10) for cooling, preferably by passing a gas or a fluid around the cavity (2), or by means of a Peltier element is provided.

26. The device according to claim 25, wherein the support (1) has a bottom (B) made of glass.

27. The device according to claim 11, wherein the seal (5) is formed by a recess 14 surrounding the lateral wall (LW) and a surrounding reinforcement (13), which is provided at the projection 4 so that it complements the recess (14) and can lock positively into the recess (14).

28. The device according to claim 27, wherein, when pressure in the cavity (2) rises, the seal (5) is self-sealing by pressing the reinforcement (13) against the recess (14).

29. A kit for preparing samples for detecting a nucleotide sequence by means of polymerase chain reaction (PCR) where the sample is contacted with an analysis solution, said kit comprising:

a) a support (1) with at least one cavity (2),

b) a lid (3) configured complementary to the cavity (2), said lid being placed onto the support (1) such that at least some of the analysis solution taken up by the cavity (2) can be displaced into a gap (G) formed between the cavity (2) and the lid (3), said lid having an electrode (6) integrally molded therein, and

c) an analysis solution comprising at least one first primer.

30. The kit according to claim 29, further comprising a second primer in the analysis solution.

31. The kit according to claim 30, wherein a third primer is bound to an internal face (I) of the lid (3) facing the cavity (2), preferably with its 5′

terminal end.