Cornell Research Foundation Inc.

University of Minnesota

Louisiana State University and Agricultural & Mechanical College

Herman Blok

Francis Barany

Robert P Hammer

George Barany

Monib Zirvi

Maria Kempe

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.

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

The present invention relates to a method of forming arrays of oligonucleotides on a solid support. This method involves providing a solid support having an array of positions each suitable for attachment of an oligonucleotide. Linkers, suitable for coupling oligonucleotides to the solid support, are attached to the solid support surface at each of the array positions. An array of a plurality of capture oligonucleotides are formed on the solid support by a series of cycles of activating selected array positions for attachment of multimer nucleotides and attaching multimer nucleotides at activated array positions. The multimer nucleotides are selected for attachment so that the capture oligonucleotides formed on the array hybridize with complementary oligonucleotide target sequences under uniform hybridization conditions.

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 supports, and the supports themselves.

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Wemeldinge, Netherlands

Herman Blok