Probes labelled with energy transfer coupled dyes

• US 5,688,648 A
• Filed: 12/19/1995
• Issued: 11/18/1997
• Est. Priority Date: 02/01/1994
• Status: Expired due to Term

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Ju, Jingyue, Mathies, Richard A., Glazer, Alexander
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): Atzel, Amy
• Application Number:
  US08/574,890
• Time in Patent Office: 700 Days
• Field of Search: 435/6, 435/77, 435/78
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• 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 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
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

Compositions are provided comprising sets of fluorescent labels carrying pairs of donor and acceptor dye molecules, designed for efficient excitation of the donors at a single wavelength and emission from the acceptor in each of the pairs at different wavelengths. The different molecules having different donor-acceptor pairs can be modified to have substantially the same mobility under separation conditions, by varying the distance between the donor and acceptor in a given pair. Particularly, the fluorescent compositions find use as labels in sequencing nucleic acids.

Claims

1. A method of identification and detection of nucleic acids in a multi-nucleic acid mixture employing detectably different fluorescent labels to detect at least two nucleic acids of interest, wherein said labels are characterized by:

(1) having a donor-acceptor fluorescent pair where said donor and said acceptor are each covalently bonded to a backbone chain at specific locations thereon with energy transfer from said donor to said acceptor; and

(2) each of the labels absorbs at substantially the same wavelength and emits at a different wavelength;

said method comprising;

covalently bonding different labels to different nucleic acids of said multi-nucleic acid mixture of form labeled nucleic acids;

detecting each of said labeled nucleic acids by irradiating at the absorption wavelength of said donor and detecting the fluorescence of each of said labels.

2. A method according to claim 1, wherein said donor absorbs light in the wavelength range of 350-800 nm and said acceptor emits light in the wavelength range of 450-1000 nm.

3. A method according to claim 2, wherein said donor-acceptor pair are 9-phenylxanthenes.