Detection of nucleic acid sequence differences using the ligase detection reaction with addressable arrays

US6852487B1 · kind B1 · utility

Assignees

• Cornell Research Foundation, Inc. · US
• BOARD OF SUPERVISORS OF LOUISIANA STATE UNIVERSITY AND AGRICULTURAL AND MECHANICAL COLLEGE · US
• Regents of the University of Minnesota · US

Inventors

• Francis Barany · Baltimore, US
• George Barany · Falcon Heights, US
• Robert P. Hammer · Baton Rouge, US
• Maria Kempe · Lund, SE
• Herman Blok · Wemeldinge, NL
• Monib Zirvi · Monmouth Junction, US

Key dates

• Filing date: Feb 4, 1997
• Grant date: Feb 8, 2005
• Expiry date: Feb 4, 2017

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C40B60/14
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detection phase. The ligation phase utilizes a ligation detection reaction between one oligonucleotide probe, which has a target sequence-specific portion and an addressable array-specific portion, and a second oligonucleotide probe, having a target sequence-specific portion and a detectable label. After the ligation phase, the capture phase is carried out by hybridizing the ligated oligonucleotide probes to a solid support with an array of immobilized capture oligonucleotides at least some of which are complementary to the addressable array-specific portion. Following completion of the capture phase, a detection phase is carried out to detect the labels of ligated oligonucleotide probes hybridized to the solid support. The ligation phase can be preceded by an amplification process. The present invention also relates to a kit for practicing this method, a method of forming arrays on solid suppor…