Method and device for preparing samples for detecting a nucleotide sequence

• US 6,440,724 B2
• Filed: 04/04/2001
• Issued: 08/27/2002
• 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, BRADLEY L
• Application Number:
  US09/825,826
• Publication Number:
  US 20010010917A1
• Time in Patent Office: 510 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 435/183, 435/287.2, 536/23.1, 536/24.33, 536/24.3
• 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

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

a) providing a support (1) defining at least one cavity, and filling the analysis solution into said at least one cavity (2),

b) placing a lid (3), which is configured complementary to a shape of the cavity (2), onto the support (1) such that at least some of the analysis solution is displaced into a gap (G) formed between the cavity (2) and the lid (3), a primer being bound to an internal face (I) of the lid (3), which extends into the cavity (2),

c) sealing the gap (G) by means of at least one seal (5, 12) provided near an opening of the cavity (2) and

d) shifting the nucleotide sequence toward the primer under the influence of an electrical field and binding the nucleotide sequence to the primer.

2. The method according to claim 1, wherein the analysis solution has a first primer added to it.

3. The method according to claim 2, wherein the analysis solution has a second primer added to it.

4. The method according to claim 3, wherein at least one of the analysis solution and primers are examined for their fluorescence properties.

5. The method according to claim 4, wherein, upon binding of the nucleotide sequence to be detected to one of the primers, a change in fluorogenic properties of the substances present in the analysis solution takes place.

6. The method according to claim 4, wherein, upon binding of the nucleotide sequence to be detected to one of the primers, a spatial relationship between two fluorophoric groups is altered such that a fluorescence reaction can be generated, altered or quenched.

7. The method according to claim 1, wherein the primer is bound to the internal face (I) of the lid (3) extending into the cavity (2), with its 5′

-terminal end.

8. The method according to claim 7, wherein the nucleotide sequence which may be present in the sample is bound to the primer.

9. The method according to claim 8, wherein after amplification cycles have been concluded, amplified nucleic acid is accumulated on the internal face (I) by binding to the primer.

10. The method according to claim 9, wherein accumulation is carried out by applying an electrical field.

11. The method according to claim 1, comprising the further step of cyclically heating and cooling the analysis solution.

12. The method according to claim 11, wherein heating is effected by means of at least one of light, a resistance heating or by passing a gas or a fluid around the cavity (2).

13. The method according to claim 11, wherein cooling is effected by passing a gas or a fluid around the cavity or by means of a Peltier element.