Title:
【発明の名称】レポーター標識鎖ターミネーター
Document Type and Number:
Japanese Patent JP2781378
Kind Code:
B2
Abstract:
A DNA sequencing system and method are described to detect the presence of radiant energy emitted from different excited reporter dye-labeled species (DNA fragments) following separation in time and/or space, and the identity of the species which emit radiant energy closely spaced in wavelength. Functions of the emitted energy are obtained which vary over the wavelengths of the closely spaced spectra in different senses and the functions ratioed, whereby the ratio is indicative of the identity of the DNA fragments. The emitting portion of the reporter-labeled DNA fragment is preferably one of a family of fluorescent dyes based on 9-carboxyethyl-6-hydroxy-3-oxo-3H-xanthene. These xanthene dyes are covalently attached to the DNA fragments through the carboxylic acid functionality, preferably via an amide linkage. The dyes may be protected by including an alkoxy group at the 9-position. A spacer may be inserted between the dye and the amine. The fluorescent dye preferably is attached to the DNA chain terminators and provides many advantages. Thus only DNA sequencing fragments resulting from bona fide termination events will carry a reporter. The DNA sequencing may also be labeled using the xanthene dyes which have general utility as fluorescent labels. Also acyclonucleoside triphosphates are described as being useful as chain terminators in DNA sequencing using a modification of the Sanger method.
Inventors:
James Merrill Prober
Ruday Joan Dam
Charles William Robertson Jr.
Frank Warden Hobbs Jr.
George Leonard Trainer
Ruday Joan Dam
Charles William Robertson Jr.
Frank Warden Hobbs Jr.
George Leonard Trainer
Application Number:
JP22153196A
Publication Date:
July 30, 1998
Filing Date:
August 22, 1996
Export Citation:
Assignee:
E.I.DU PONT DE NEMOURS AND COMPANY
International Classes:
C07D311/84; C07D311/86; C07D405/04; C07D405/12; C07D405/14; C07D407/12; C07D471/04; C07D487/04; C07D311/82; C07F9/6512; C07F9/655; C07F9/6558; C07F9/6561; C07H19/06; C07H19/10; C07H19/14; C07H19/20; C07H21/00; C09B11/28; C09K11/06; C12N15/09; C12Q1/68; G01N27/447; G01N33/50; G01N33/58; (IPC1-7): C07F9/655; C07F9/6561; C07H19/10; C07H19/14
Attorney, Agent or Firm:
Yoshikazu Tani (2 outside)