Dyes and labeled molecules

• US 8,530,195 B2
• Filed: 06/22/2010
• Issued: 09/10/2013
• Est. Priority Date: 03/17/2005
• Status: Active Grant

• Current Assignee: Allelogic Biosciences Corporation, Biotium, Inc.
• Original Assignee: Biotium, Inc.
• Inventors: Mao, Fei, Leung, Wai-Yee, Xin, Xing
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US12/820,983
• Publication Number:
  US 20100317016A1
• Time in Patent Office: 1,176 Days
• Field of Search: 435/6.1, 435/91.1, 536/26.6
• US Class Current: 435/91.1
• CPC Class Codes: C07D 209/08 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
• CPC Class Codes: C07D 213/38 having only hydrogen or hydrocarbon radicals attached to the substituent nitrogen atom
• CPC Class Codes: C07D 221/10 Aza-phenanthrenes
• CPC Class Codes: C07D 417/06 linked by a carbon chain containing only aliphatic carbon atoms
• CPC Class Codes: C07D 417/14 containing three or more hetero rings
• CPC Class Codes: C07D 491/147 the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C09B 15/00 Acridine dyes
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2563/173 staining/intercalating agent, e.g. ethidium bromide

Abstract

Dimeric and trimeric nucleic acid dyes, and associated systems and methods are provided. Such a dye may form a hairpin-like structure that enables it to stain nucleic acids via a release-on-demand mechanism, for example. Such a dye may have low background fluorescence in the absence of nucleic acids and high fluorescence in the presence of nucleic acids, upon binding therewith, for example. A dye provided herein may be useful in a variety of applications, such as in DNA quantitation in real-time PCR, for example.

Claims

1. A method of performing a nucleic acid amplification reaction comprising:

(i) providing a reaction mixture comprising said sample and reagents necessary for amplifying nucleic acid;

(ii) subjecting said reaction mixture to a polymerization reaction under conditions suitable for formation of amplified nucleic acid;

(iii) adding a fluorescent nucleic acid dye, wherein said addition takes place before, during, or after step (ii), and wherein said dye has the formula;

2. The method of claim 1, wherein the fluorescent nucleic acid dye has the structure:

3. The method of claim 1, wherein T is a substituted nitrogen.

4. The method of claim 1, wherein each of Q₁ and Q₂ is independently represented by the structure:

5. The method of claim 1, wherein each of Q1 and Q2 is independently represented by the structure:

6. The method of claim 1, wherein each of Q1 and Q2 is independently represented by the structure:

7. The method of claim 1, wherein each of Q1 and Q2 is independently represented by the structure: