Solid-phase reactions

• US 20030119020A1
• Filed: 07/17/2002
• Published: 06/26/2003
• Est. Priority Date: 07/17/2001
• Status: Active Grant

• Current Assignee: Third Wave Technologies Incorporated (Hologic Incorporated)
• Original Assignee: Northwestern University
• Inventors: Kelso, David, Neri, Bruce P., Lyamichev, Victor, Kwiatkowski, Robert W. Jr., Hall, Jeff, Stevens, Priscilla Wilkins, Smith, Lloyd
• Granted Patent: US 7,485,443 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/683 involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
• CPC Class Codes: C12Q 2561/109 Invader technology
• CPC Class Codes: C12Q 2565/549 characterised by the capture oligonucleotide being a reporter labelled capture oligonucleotide

Abstract

The present invention relates to systems, compositions, and methods for the detection and characterization of nucleic acid sequences and variations in nucleic acid sequences. The present invention relates to methods for attaching nucleic acids to solid supports and modifying nucleic acids. For example, in some embodiments, the 5′ nuclease activity of a cleavage agent is used to cleave a cleavage structure formed on the solid support, the occurrence of the cleavage event indicating the presence of specific nucleic acid sequences.

Claims

1. A method for detecting polymorphisms in unamplified DNA, comprising:

a. providing;

i. unamplified DNA;

ii. reagents for conducting an invasive cleavage reaction, wherein at least one of said reagents is attached to a solid surface;

b. treating said unamplified DNA with said reagents under conditions such that the presence or absence of a polymorphism in said genomic DNA is identified.

2. The method of claim 1, wherein said reagents are configured to detect a plurality of different polymorphisms in said unamplified DNA.

3. The method of claim 2, wherein said plurality of different polymorphisms comprises at least 100 different polymorphisms.

4. The method of claim 2, wherein said plurality of different polymorphisms comprises at least 1000 different polymorphisms.

5. The method of claim 2, wherein said plurality of different polymorphisms comprises at least 1,000,000 different polymorphisms.

6. The method of claim 1, wherein said solid surface comprises a microsphere.

7. The method of claim 1, wherein said solid surface comprises a plurality of microspheres.

8. The method of claim 1, wherein said treating step is carried out in a single reaction vessel.

9. The method of claim 2, wherein said unamplified DNA comprises 50 micrograms or less of genomic DNA.

10. The method of claim 1, wherein said unamplified DNA comprises genomic DNA.

11. A system for detecting polymorphisms in unamplified DNA, comprising:

reagents for conducting an invasive cleavage reaction, wherein at least one of said reagents is attached to a microsphere solid surface.

12. The system of claim 11, wherein said reagents comprise reagents for detecting a plurality of polymorphisms.

13. The system of claim 12, wherein said plurality of different polymorphisms comprises at least 100 different polymorphisms.

14. The system of claim 12, wherein said plurality of different polymorphisms comprises at least 1000 different polymorphisms.

15. The system of claim 12, wherein said plurality of different polymorphisms comprises at least 1,000,000 different polymorphisms.

16. The system of claim 11, wherein said unamplified DNA comprises genomic DNA.