Compounds and methods for labeling oligonucleotides

• US 7,476,735 B2
• Filed: 05/22/2006
• Issued: 01/13/2009
• Est. Priority Date: 05/20/2005
• Status: Active Grant

• Current Assignee: Integrated DNA Technologies (Danaher Corporation)
• Original Assignee: Integrated DNA Technologies (Danaher Corporation)
• Inventors: Yong, Yawful, Laikhter, Andrei, Walder, Joseph A., Behlke, Mark, Podyminogin, Mikhail
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US11/438,606
• Publication Number:
  US 20060263816A1
• Time in Patent Office: 967 Days
• Field of Search: 435/6, 536/23.1, 536/24.3, 536/25.32, 536/26.6, 534/558, 530/391.3
• US Class Current: 536/26.6
• CPC Class Codes: C07C 245/10 with nitrogen atoms of azo groups bound to carbon atoms of six-membered aromatic rings being part of condensed ring systems
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer

Abstract

The invention provides a novel method of labeling oligonucleotides, with reporter moieties, including but not limited to, quenchers, fluorophores, biotin, digoxigenin, peptides and proteins. In addition, this invention provides a method of detecting hybridization of oligonucleotides. This invention also provides novel azo quenchers having the general formula shown below.

The invention further provides compositions comprising labeled oligonucleotides and solid supports. The invention also provides kits comprising at least one composition of the present invention.

Claims

1. A method for linking a reporter moiety to an oligonucleotide comprising reacting a reporter moiety having an oxime forming nucleophile substituent with an oxo substituted reactant coupled to a solid support to form an oxime bond between the reporter moiety and the reactant.

2. The method of claim 1, wherein the reporter moiety is selected from the group consisting of quenchers, fluorophores, biotin, digoxigenin, peptides, proteins, antibodies, glycoproteins and sugars.

3. The method of claim 1, wherein the reporter moiety is a compound of Formula (I)

4. The method of claim 3, wherein the oxime forming nucleophile is aminooxy.

5. The method of claim 3, wherein the oxime forming nucleophile is hydrazine.

6. The method of claim 1, wherein the reporter moiety is

7. The method of claim 1, wherein more than one reporter moiety is linked to the labeled oligonucleotide, and wherein the reporter moieties are the same or different.

8. The method of claim 1, wherein the reporter moiety is selected from the group consisting of compound 34, compound 35, compound 36 and compound 37.