IP Library › Granted Patent US 7,601,496
Granted Patent B2
US 7,601,496 · App. 11/198,746 · Granted Oct 13, 2009

Cleavage of nucleic acids

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Quick Facts
Patent No.
US 7,601,496
App. No.
11/198,746
Granted
Oct 13, 2009
Kind
B2
Abstract

The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5′ nucleases and 3′ exonucleases, are used to detect and identify nucleic acids derived from microorganisms. Methods are provided which allow for the detection and identification of bacterial and viral pathogens in a sample.

Claims (17)

1. A method for characterizing a target nucleic acid molecule comprising: a) exposing a sample comprising a target nucleic acid molecule to an isolated polymerase lacking 5′nuclease activity to form an amplified product, b) forming a nucleic acid complex by annealing a second nucleic acid molecule to said amplified product, c) cleaving said nucleic acid complex with an isolated flap endonuclease, wherein said isolated flap endonuclease is selected from the group consisting of murine FEN-1 endonuclease and calf 5′to 3′ exonuclease; and d) detecting the cleavage of said nucleic acid complex.

2. The method of claim 1 , wherein said target nucleic acid molecule is DNA.

3. The method of claim 1 , wherein said target nucleic acid molecule is RNA.

4. The method of claim 1 , wherein said target nucleic acid is cDNA.

5. The method of claim 1 , wherein said target nucleic acid molecule is from a pathogen.

6. The method of claim 1 , wherein said target nucleic acid is synthetic.

7. The method of claim 1 , wherein said amplified product is produced by polymerase chain reaction.

8. The method of claim 5 , wherein said pathogen is a virus.

9. The method of claim 5 , wherein said pathogen is a bacterium.

10. The method of claim 1 , wherein said second nucleic acid molecule is attached to a solid support.

11. The method of claim 1 , wherein said second nucleic acid molecule comprises a label.

12. The method of claim 11 , wherein said label comprises a fluorescent label.

13. The method of claim 1 , wherein said isolated polymerase lacking 5′nuclease activity is Taq DNA polymerase Stoffel fragment.

14. The method of claim 1 , wherein said isolated polymerase lacking 5′nuclease activity is Pfu DNA polymerase.

15. The method of claim 1 , wherein said isolated polymerase lacking 5′nuclease activity is Tli DNA polymerase.

16. The method of claim 1 , wherein said amplified product comprises a hairpin structure.

17. The method of claim 1 , wherein said nucleic acid complex comprises single stranded and double stranded regions.

Assignments (8)
RELEASE OF SECURITY INTEREST Recorded Apr 28, 2026
From: BANK OF AMERICA, N.A., AS COLLATERAL AGENT
To: HOLOGIC, INC., ON ITS OWN BEHALF AND AS SUCCESSOR-BY-MERGER TO DIRECT RADIOGRAPHY CORP.; CYTYC CORPORATION, ON ITS OWN BEHALF AND AS SUCCESSOR-BY-MERGER TO BIOLUCENT, LLC; CYTYC SURGICAL PRODUCTS, LLC, AS SUCCESSOR-BY-CONVERSION TO CYTYC SURGICAL PRODUCTS, LIMITED PARTNERSHIP; GEN-PROBE INCORPORATED, ON ITS OWN BEHALF AND AS SUCCESSOR-BY-MERGER TO THIRD WAVE TECHNOLOGIES, INC.; GEN-PROBE PRODESSE, INC.; SUROS SURGICAL SYSTEMS, INC.
Reel/Frame 075566/0039 →
CORRECTIVE ASSIGNMENT TO CORRECT THE INCORRECT PATENT NO. 8081301 PREVIOUSLY RECORDED AT REEL: 028810 FRAME: 0745. ASSIGNOR(S) HEREBY CONFIRMS THE SECURITY AGREEMENT. Recorded Nov 9, 2017
From: HOLOGIC, INC.; BIOLUCENT, LLC; CYTYC CORPORATION; CYTYC SURGICAL PRODUCTS, LIMITED PARTNERSHIP; SUROS SURGICAL SYSTEMS, INC.; THIRD WAVE TECHNOLOGIES, INC.; GEN-PROBE INCORPORATED
To: GOLDMAN SACHS BANK USA
Reel/Frame 044432/0565 →
CORRECTIVE ASSIGNMENT TO CORRECT THE INCORRECT PATENT NO. 8081301 PREVIOUSLY RECORDED AT REEL: 035820 FRAME: 0239. ASSIGNOR(S) HEREBY CONFIRMS THE SECURITY INTEREST RELEASE. Recorded Nov 9, 2017
From: GOLDMAN SACHS BANK USA, AS COLLATERAL AGENT
To: HOLOGIC, INC.; BIOLUCENT, LLC; CYTYC CORPORATION; CYTYC SURGICAL PRODUCTS, LIMITED PARTNERSHIP; SUROS SURGICAL SYSTEMS, INC.; THIRD WAVE TECHNOLOGIES, INC.; GEN-PROBE INCORPORATED
Reel/Frame 044727/0529 →
SECURITY AGREEMENT Recorded Aug 7, 2015
From: HOLOGIC, INC.; BIOLUCENT, LLC; CYTYC CORPORATION; CYTYC SURGICAL PRODUCTS, LIMITED PARTNERSHIP; DIRECT RADIOGRAPHY CORP.; GEN-PROBE INCORPORATED; GEN-PROBE PRODESSE, INC.; SUROS SURGICAL SYSTEMS, INC.; THIRD WAVE TECHNOLOGIES, INC.
To: BANK OF AMERICA, N.A., AS COLLATERAL AGENT
Reel/Frame 036307/0199 →
SECURITY INTEREST RELEASE REEL/FRAME 028810/0745 Recorded Jun 4, 2015
From: GOLDMAN SACHS BANK USA, AS COLLATERAL AGENT
To: HOLOGIC, INC.; BIOLUCENT, LLC; CYTYC CORPORATION; CYTYC SURGICAL PRODUCTS, LIMITED PARTNERSHIP; SUROS SURGICAL SYSTEMS, INC.; THIRD WAVE TECHNOLOGIES, INC.; GEN-PROBE INCORPORATED
Reel/Frame 035820/0239 →
SECURITY AGREEMENT Recorded Aug 1, 2012
From: HOLOGIC, INC.; BIOLUCENT, LLC; CYTYC CORPORATION; CYTYC SURGICAL PRODUCTS, LIMITED PARTNERSHIP; SUROS SURGICAL SYSTEMS, INC.; THIRD WAVE TECHNOLOGIES, INC.; GEN-PROBE INCORPORATED
To: GOLDMAN SACHS BANK USA
Reel/Frame 028810/0745 →
TERMINATION OF PATENT SECURITY AGREEMENTS AND RELEASE OF SECURITY INTERESTS Recorded Aug 26, 2010
From: GOLDMAN SACHS CREDIT PARTNERS, L.P., AS COLLATERAL AGENT
To: HOLOGIC, INC.; R2 TECHNOLOGY, INC.; SUROS SURGICAL SYSTEMS, INC.; BIOLUCENT, LLC; DIRECT RADIOGRAPHY CORP.; CYTYC SURGICAL PRODUCTS II LIMITED PARTNERSHIP; CYTYC SURGICAL PRODUCTS LIMITED PARTNERSHIP; CYTYC CORPORATION; CYTYC SURGICAL PRODUCTS III, INC.; CYTYC PRENATAL PRODUCTS CORP.; THIRD WAVE TECHNOLOGIES, INC.
Reel/Frame 024892/0001 →
SECURITY AGREEMENT Recorded Jul 29, 2008
From: THIRD WAVE TECHNOLOGIES, INC.
To: GOLDMAN SACHS CREDIT PARTNERS L.P., AS COLLATERAL AGENT
Reel/Frame 021301/0780 →