RNase H-Based Assays Utilizing Modified RNA Monomers

• US 20140106433A1
• Filed: 08/05/2013
• Published: 04/17/2014
• Est. Priority Date: 04/30/2008
• Status: Active Grant

• Current Assignee: Integrated DNA Technologies (Danaher Corporation)
• Original Assignee: Integrated DNA Technologies (Danaher Corporation)
• Inventors: Walder, Joseph Alan, Behlke, Mark Aaron, Rose, Scott, Dobosy, Joesph
• Granted Patent: US 9,644,198 B2
• Time in Patent Office: Days
• US Class Current: 435/188
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/6853 using modified primers or templates
• CPC Class Codes: C12Q 2521/301 Endonuclease
• CPC Class Codes: C12Q 2525/121 incorporating both deoxyribonucleotides and ribonucleotides
• CPC Class Codes: C12Q 2525/186 incorporating a non-extendable or blocking moiety
• CPC Class Codes: C12Q 2549/101 Hot start

Abstract

The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3′ end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.

Claims

1. A hot start enzyme composition comprising an enzyme and a chemical modification, wherein the chemical modification inactivates the enzyme.

2. The hot start enzyme composition of claim 1 wherein the enzyme is Pyrococcus abysii.

3. The hot start enzyme composition of claim 2 wherein the Pyrococcus abysii enzyme comprises the polypeptide of SEQ ID NO. 4.

4. The hot start enzyme composition of claim 1 wherein the chemical modification is a maleic acid anhydride analog.

5. The hot start enzyme composition of claim 4 wherein the maleic acid anhydride analog is citroconic anhydride.

6. A hot start enzyme composition comprising an enzyme and an antibody bound to the enzyme, wherein the antibody inactivates the enzyme at low temperatures.

7. The hot start enzyme composition of claim 6 wherein the enzyme is Pyrococcus abysii.

8. The hot start enzyme composition of claim 7 wherein the Pyrococcus abysii enzyme comprises the polypeptide of SEQ ID NO. 4.

9. The hot start enzyme composition of claim 7 wherein the antibody is an antibody that specifically binds Pyrococcus abysii