Probes labeled with energy transfer coupled dyes

• US 20060024743A1
• Filed: 09/29/2005
• Published: 02/02/2006
• Est. Priority Date: 02/01/1994
• Status: Abandoned Application

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Mathies, Richard, Ju, Jingyue, Glazer, Alexander
• Application Number:
  US11/241,121
• Publication Number:
  US 20060024743A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2535/101 Sanger sequencing method, i.e. oligonucleotide sequencing using primer elongation and dideoxynucleotides as chain terminators
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: C12Q 2565/1015 labels being on the same oligonucleotide
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

Fluorescent labels having at least one donor and at least one acceptor fluorophore bonded to a polymeric backbone in energy transfer relationship, as well as methods for their use, are provided. Of particular interest are the subject labels wherein the polymeric backbone is a nucleic acid and the donor fluorophore is bonded to the 5′ terminus of said nucleic acid. Such labels find use as primers in applications involving nucleic acid chain extension, such as sequencing, PCR and the like.

Claims

1-29. -29. (canceled)

30. A kit for use in simultaneously detecting a plurality of components, said kit comprising a set of labels consisting essentially of at least four labels, each of said labels having essentially only one specific chemical structure, each of said labels comprising a donor-acceptor fluorescent pair wherein said donor and said acceptor are covalently bonded to a covalently bonded chain of atoms such that there is energy transfer to said acceptor, and each of said labels absorptive of light at substantially the same wavelength and emitting light at a different wavelength.

31. A kit for use in simultaneously detecting a plurality of components, said kit comprising a set of labels consisting essentially of at least four fluorescent energy-transfer labels,

provided that said first fluorescent energy-transfer label comprises a first fluorescent donor, a first fluorescent acceptor, and a first covalently bonded chain of atoms, said second fluorescent energy-transfer label comprises a second fluorescent donor, a second fluorescent acceptor, and a second covalently bonded chain of atoms, said third fluorescent energy-transfer label comprises a third fluorescent donor, a third fluorescent acceptor, and a third covalently bonded chain of atoms, and said fourth fluorescent energy-transfer label comprises a fourth fluorescent donor, a fourth fluorescent acceptor, and a fourth covalently bonded chain of atoms,

further provided that said first donor and said first acceptor are covalently bonded to said first covalently bonded chain of atoms to produce a first fluorescent energy-transfer label having essentially only one specific chemical structure with energy transfer from said first donor to said first acceptor, said second donor and said second acceptor are covalently bonded to said second covalently bonded chain of atoms to produce a second fluorescent energy-transfer label having essentially only one specific chemical structure with energy transfer from said second donor to said second acceptor, said third donor and said third acceptor are covalently bonded to said third covalently bonded chain of atoms to produce a third fluorescent energy-transfer label having essentially only one specific chemical structure with energy transfer from said third donor to said third acceptor, and said fourth donor and said fourth acceptor are covalently bonded to said fourth covalently bonded chain of atoms to produce a fourth fluorescent energy-transfer label having essentially only one specific chemical structure with energy transfer from said fourth donor to said fourth acceptor, and

provided further that the absorption wavelengths of said first donor, said second donor, said third donor, and said fourth donor are substantially the same and said first acceptor, said second acceptor, said third acceptor, and said fourth acceptor, each have different emission wavelengths.