DDAO compounds as fluorescent reference standards

Patent metadata

Field	Value
Publication number	US-10131936-B2
Application number	US-201715707157-A
Country	US
Kind code	B2
Filing date	Sep 18, 2017
Priority date	Dec 29, 2010
Publication date	Nov 20, 2018
Grant date	Nov 20, 2018

Abstract

According to the present teachings, methods and compositions are provided that utilize at least one reference dye of formula (I):

In some embodiments, a method comprises measuring a detection signal of a reporter dye and at least one reference dye of formula (I). In some embodiments, a composition comprises a reference dye of formula (1), a buffer, a selection of nucleotides and a protein.

First claim

What is claimed is:

1. A kit for amplification comprising:
a buffer,
a selection of nucleotides,
a reference dye, and
instructions for carrying out an assay;
wherein the reference dye has a structure according to formula (I):

wherein:
each of R 1 to R 3 and R 6 to R 8 is independently —H, halogen, —CO 2 H, —CO 2 R, —SO 3 H, —SO 3 R, —CH 2 CO 2 H, —CH 2 CO 2 R, —CH 2 SO 3 H, —CH 2 SO 3 R, —CH 2 NH 2 , —CH 2 NHR, —NO 2 , C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, and substituted C 1 -C 6 alkoxy, wherein R is C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, and substituted C 1 -C 6 alkoxy;
R 4 and R 5 taken separately are selected from C 1 -C 6 alkyl, and C 1 -C 6 substituted alkyl, or R 4 and R 5 taken together are selected from C 3 -C 7 cycloalkyl, C 4 -C 7 unsaturated cycloalkyl, C 3 -C 7 substituted cycloalkyl, or C 4 -C 7 substituted unsaturated cycloalkyl.

2. The kit of claim 1 , wherein the buffer is selected from tris, tricene, bicene, HEPES, and MOPS.

3. The kit of claim 1 , wherein the buffer further comprises a chloride salt selected from potassium, lithium, magnesium and manganese, and a surfactant.

4. The kit of claim 3 , wherein the surfactant is selected from a polysorbate surfactant, a polyoxyethylene surfactant, a polyethoxylated phenol surfactant, and a zwitterionic surfactant.

5. The kit of claim 1 , wherein the selection of nucleotides is from deoxynucloetides (dNPTs).

6. The kit of claim 5 , wherein the selection of nucleotides is further from dideoxynucleotides (didNTPs), deaza-GTP, deaza-dGTP, and 2′-deoxyinosine 5′-triphosphate (dITP).

7. The kit of claim 1 , further comprising primers.

8. The kit of claim 1 , wherein the kit components are lyophilized.

9. The kit of claim 1 , wherein the kit further comprises at least one protein moiety.

10. The kit of claim 1 , wherein the contents are combined with at least one protein moiety.

11. The kit of claim 9 , wherein the at least one protein moiety is selected from a DNA polymerase, a ligase, a reverse transcriptase, a ribonuclease, a glycosylase, a single strand binding protein, a pyrophosphatase, an albumin, and a gelatin.

12. The kit of claim 10 , wherein the at least one protein moiety is selected from a DNA polymerase, a ligase, a reverse transcriptase, a ribonuclease, a glycosylase, a single strand binding protein, a pyrophosphatase, an albumin, and a gelatin.

Assignees

• Life Technologies Corp

Inventors

• Benson Scott C
• Monighetti Cinna
• Koepf Sandy M

Classifications

• C12Q2561/113

  Real time assay · CPC title

• C12Q1/6818 Primary

  involving interaction of two or more labels, e.g. resonant energy transfer · CPC title

• C09B15/00 Primary

  Acridine dyes · CPC title

• C12Q2527/125

  Specific component of sample, medium or buffer · CPC title

• C12Q2545/101

  with an internal standard/control · CPC title