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

摘要

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.

主权项

1. A method comprising:
providing a collection of oligonucleotides wherein each type of oligonucleotide is greater than 16 nucleotides and differs in nucleotide sequence, when aligned to another type of oligonucleotide in the collection, by at least 25%; providing a sample comprising a plurality of composite oligonucleotides, each composite oligonucleotide comprising (i) a zip-code portion, and (ii) one or more further nucleotide sequences; contacting the sample comprising the plurality of composite oligonucleotides with the collection of oligonucleotides under conditions effective to hybridize the zip-code portion of each composite oligonucleotide to its complementary oligonucleotide in the collection; and detecting the one or more of the plurality of composite oligonucleotides hybridized to their complementary oligonucleotides in the collection.