DDAO compounds as fluorescent reference standards

• US 10,131,936 B2
• Filed: 09/18/2017
• Issued: 11/20/2018
• Est. Priority Date: 12/29/2010
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Benson, Scott C., Monighetti, Cinna, Koepf, Sandy M.
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US15/707,157
• Publication Number:
  US 20180127811A1
• Time in Patent Office: 428 Days
• Field of Search: 435 61, 435 911
• CPC Class Codes: C09B 15/00 Acridine dyes
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2545/101 with an internal standard/control
• CPC Class Codes: C12Q 2561/113 Real time assay
• CPC Class Codes: C12Q 2563/107 fluorescence

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.

Claims

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);

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.