Fluorescent dye compounds, conjugates and uses thereof

• US 9,745,336 B2
• Filed: 03/28/2014
• Issued: 08/29/2017
• Est. Priority Date: 09/16/2004
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Graham, Ronald, Zou, Ruiming, Upadhya, Krishna, Benson, Scott
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US14/229,663
• Publication Number:
  US 20140303003A1
• Time in Patent Office: 1,250 Days
• Field of Search: 536 266
• CPC Class Codes: C07D 219/06 Oxygen atoms
• 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: C09K 11/06 containing organic luminescent materials
• CPC Class Codes: C09K 2211/1011 Condensed systems
• CPC Class Codes: C09K 2211/1029 containing one nitrogen atom as the heteroatom
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The present teachings generally relate to fluorescent dyes, linkable forms of fluorescent dyes, energy transfer dyes, reagents labeled with fluorescent dyes and uses thereof.

Claims

1. A composition comprising:

a) an oligonucleotide comprising a fluorescent dye; and

b) an internal standard dye compound;

wherein the fluorescent dye of the oligonucleotide and the internal standard dye compound are spectrally resolvable and further wherein the internal standard dye compound is selected from;

2. The composition of claim 1, wherein the internal standard dye compound is selected from compounds of formula (I).

3. The composition of claim 1, wherein R₆ and R₇ are each independently halogens selected from fluorine, chlorine, and bromine.

4. The composition of claim 1, wherein R₆ and R₇ are fluorine or R₆ and R₇ are chlorine.

5. The composition of claim 1, wherein the internal dye standard compound is selected from compounds of formula (I) and R₂ is selected from —

SO₃H, —

SO₃R, —

CH₂NH₂, —

CH₂NHR, and —

NO₂.

6. A kit comprising:

a) an oligonucleotide labeled with a fluorescent dye; and

b) an internal standard dye compound;

wherein the fluorescent dye of the oligonucleotide and the internal standard dye compound are spectrally resolvable and further wherein the internal standard dye compound is selected from;

7. The kit of claim 6, wherein the internal standard dye compound is selected from compounds having a structure of formula (I).

8. The kit of claim 7, wherein R₆ and R₇ are each independently halogens selected from fluorine, chlorine, and bromine.

9. The kit of claim 8, wherein R₆ and R₇ are fluorine or R₆ and R₇ are chlorine.

10. The kit of claim 9, wherein the internal dye standard compound is selected from compounds of formula (I) and R₂ is selected from —

SO₃H, —

SO₃R, —

CH₂NH₂, —

CH₂NHR, and —

NO₂.

11. A composition comprising:

a) an oligonucleotide comprising a fluorescent dye; and

b) an internal standard dye compound;

wherein the fluorescent dye of the oligonucleotide and the internal standard dye compound are spectrally resolvable and further wherein the internal standard dye compound is;

12. The composition of claim 11, wherein R₆ and R₇ are each independently halogens selected from fluorine, chlorine, and bromine.

13. The composition of claim 12, wherein R₆ and R₇ are fluorine or R₆ and R₇ are chlorine.

14. The composition of claim 11, wherein R₂ is selected from —

SO₃H, —

SO₃R, —

CH₂NH₂, —

CH₂NHR, and —

NO₂.

15. A kit comprising:

a) an oligonucleotide labeled with a fluorescent dye; and

b) an internal standard dye compound;

wherein the fluorescent dye of the oligonucleotide and the internal standard dye compound are spectrally resolvable and further wherein the internal standard dye compound is;

16. The kit of claim 15, wherein R₆ and R₇ are each independently halogens selected from fluorine, chlorine, and bromine.

17. The kit of claim 16, wherein R₆ and R₇ are fluorine or R₆ and R₇ are chlorine.

18. The kit of claim 15, wherein R₂ is selected from —

SO₃H, —

SO₃R, —

CH₂NH₂, —

CH₂NHR, and —

NO₂.