Method for DNA sequencing and gene identification

• US 20030143549A1
• Filed: 01/29/2002
• Published: 07/31/2003
• Est. Priority Date: 01/29/2002
• Status: Abandoned Application

• Current Assignee: Eastman Kodak Company
• Original Assignee: Eastman Kodak Company
• Inventors: Yang, Zhihao, Qiao, Tiecheng A.
• Application Number:
  US10/059,988
• Publication Number:
  US 20030143549A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/155 Particles of a defined size, e.g. nanoparticles

Abstract

A method for single molecule identification of a target DNA molecule in a random coil state having the following steps: a) attaching an optically distinguishable material to a DNA sequence recognition unit; b) hybridizing the DNA sequence recognition unit to the target DNA molecule in a random coil state to form a hybridized DNA complex in a random coil state; c) stretching the hybridized DNA complex in a random coil state to form a hybridized DNA complex in a substantially linear configuration; and d) detecting the optically distinguishable material in a sequential manner along the substantially linear hybridized DNA complex, thereby identifying the target DNA molecule.

Claims

1. A method for single molecule identification of a target DNA molecule in a random coil state comprising the following steps:

a) attaching an optically distinguishable material to a DNA sequence recognition unit;

b) hybridizing said DNA sequence recognition unit to said target DNA molecule in a random coil state to form a hybridized DNA complex in a random coil state;

c) stretching said hybridized DNA complex in a random coil state to form a hybridized DNA complex in a substantially linear configuration; and

d) detecting said optically distinguishable material in a sequential manner along said substantially linear hybridized DNA complex, thereby identifying said target DNA molecule.

2. The method of claim 1 wherein said optically distinguishable material comprises colored microparticles.

3. The method of claim 1 wherein said optically distinguishable material comprises microparticles having different shapes.

4. The method of claim 2 wherein said colored microparticles comprise dyes, dye aggregates, pigments or nanocrystals.

5. The method of claim 1 wherein said DNA sequence recognition unit comprises DNA, DNA fragments, synthetic oligonucleotides or peptide nucleic acids.

6. The method of claim 1 wherein said DNA sequence recognition units comprise any protein scaffold or synthetic molecular moiety capable of recognizing a specific DNA sequence.

7. The method of claim 1 wherein said stretching of said hybridized DNA complex in a random coil state to form a hybridized DNA complex in a substantially linear configuration is accomplished by using a mechanical means.

8. A method for single molecule identification of a target DNA molecule in a random coil state comprising the following steps:

a) stretching said target DNA molecule in a random coil state to form a substantially linear configuration;

b) attaching an optically distinguishable material to a DNA sequence recognition unit;

c) hybridizing said DNA sequence recognition unit to said target DNA molecule in a substantially linear configuration to form a hybridized DNA complex in a substantially linear configuration; and

d) detecting said optically distinguishable material in a sequential manner along said substantially linear hybridized DNA complex, thereby identifying said target DNA molecule.

9. The method of claim 8 wherein said optically distinguishable material comprises colored microparticles.

10. The method of claim 8 wherein said optically distinguishable material comprises microparticles having different shapes.

11. The method of claim 9 wherein said colored microparticles comprise dyes, dye aggregates, pigments or nanocrystals.

12. The method of claim 8 wherein said DNA sequence recognition unit comprises DNA, DNA fragments, synthetic oligonucleotides or peptide nucleic acids.

13. The method of claim 8 wherein said DNA sequence recognition units comprise any protein scaffold or synthetic molecular moiety capable of recognizing a specific DNA sequence.

14. The method of claim 8 wherein said stretching of said hybridized DNA complex in a random coil state to form a hybridized DNA complex in a substantially linear configuration is accomplished by using a mechanical means.