Method of analyzing double stranded DNA

• US 6,924,105 B2
• Filed: 10/24/2001
• Issued: 08/02/2005
• Est. Priority Date: 10/25/2000
• Status: Expired due to Fees

• Current Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Sudo, Yukio, Makino, Yoshihiko
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): LU, FRANK WEI MIN
• Application Number:
  US10/039,642
• Publication Number:
  US 20020177146A1
• Time in Patent Office: 1,378 Days
• Field of Search: 435/6, 435/7.1, 435/91.1, 435/183, 436/94, 436/501, 536/23.1, 536/24.3, 536/24.33, 536/25.3, 204/450, 424/130.1, 530/300, 530/350
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2522/101 Single or double stranded nucleic acid binding proteins
• CPC Class Codes: C12Q 2563/173 staining/intercalating agent, e.g. ethidium bromide
• CPC Class Codes: C12Q 2565/125 Electrophoretic separation
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites

Abstract

The object of the present invention is to provide a method for analyzing double stranded DNA directly without denaturation. According to the present invention, there is provided a method of analyzing double stranded DNA present in an analyte, which comprises the steps of:

• (1) contacting the analyte with a double stranded DNA recognizing substance immobilized on a support, and
• (2) measuring double stranded DNA bound to the double stranded DNA recognizing substance.

Claims

1. A method of analyzing a double stranded DNA present in an analyte, which comprises the steps of:

(1) contacting the analyte with a double stranded DNA recognizing substance immobilized on a support,

(2) applying an electric field between the support on which the double stranded DNA recognizing substance is immobilized and the analyte, to direct the double stranded DNA present in the analyte toward the double stranded DNA recognizing substance immobilized on the support,

(3) applying an electric field in a direction which is opposite to the direction of the electric field applied in step (2), and

(4) measuring the double stranded DNA bound to the double stranded DNA recognizing substance,

wherein the support and the analyte is within the electric field.

2. The method of analysis according to claim 1 wherein the double stranded DNA recognizing substance is a double stranded DNA recognizing antibody.

3. The method of analysis according to claim 1 wherein the double stranded DNA recognizing substance is a DNA transcription factor.

4. The method of analysis according to claim 1 wherein the double stranded DNA recognizing substance is a protein having Zn finger motif or Ring finger motif.

5. The method of analysis according to claim 1 wherein the double stranded DNA recognizing substance is a peptide nucleic acid.

6. The method of analysis according to claim 1 wherein, in the step of measuring the double stranded DNA bound to the double stranded DNA recognizing substance, an insertion agent which recognizes double stranded DNA is added to a reaction system, and the double stranded DNA present in the analyte is measured by detecting the insertion agent inserted into the double stranded DNA.

7. The method of analysis according to claim 6 wherein the insertion agent is a DNA intercalator.

8. The method of analysis according to claim 7 wherein the DNA intercalator has an electrochemical activity, and the double stranded DNA present in the analyte is measured by detecting the DNA intercalator by electrochemical means.

9. The method of analysis according to claim 7 wherein the DNA intercalator is detected by a fluorescence, luminescence or surface plasmon method.