DETECTION OF NUCLEIC ACID SEQUENCE DIFFERENCES USING THE LIGASE DETECTION REACTION WITH ADDRESSABLE ARRAYS
First Claim
1. A kit for identifying one or more of a plurality of sequences differing by single-base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences comprising:
- a ligase;
a collection of oligonucleotide probe sets, each set characterized by (a) a first oligonucleotide probe comprising a target-specific portion and (b) a second oligonucleotide probe comprising a target-specific portion and a further portion, wherein the further portion comprises a nucleic acid sequence of greater than sixteen nucleotides that differs for each different target-specific portion, and wherein the nucleic acid sequence of a complement to one further portion differs substantially from the nucleic acid sequence of a complement to another further portion in the collection, when aligned to each other.
1 Assignment
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Accused Products
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.
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Citations
11 Claims
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1. A kit for identifying one or more of a plurality of sequences differing by single-base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences comprising:
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a ligase; a collection of oligonucleotide probe sets, each set characterized by (a) a first oligonucleotide probe comprising a target-specific portion and (b) a second oligonucleotide probe comprising a target-specific portion and a further portion, wherein the further portion comprises a nucleic acid sequence of greater than sixteen nucleotides that differs for each different target-specific portion, and wherein the nucleic acid sequence of a complement to one further portion differs substantially from the nucleic acid sequence of a complement to another further portion in the collection, when aligned to each other. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A kit for identifying one or more of a plurality of target nucleotide sequences in a sample comprising:
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a ligase; a plurality of oligonucleotide probe sets, each set characterized by (a) a first oligonucleotide probe, having a target-specific portion and an address-specific portion and (b) a second oligonucleotide probe, having a target-specific portion; and a collection of capture oligonucleotides wherein each type of capture oligonucleotide in the collection comprises a nucleotide sequence complementary to an address-specific portion, wherein the address-specific portion is comprised of a nucleotide sequence which is distinct from that of the target-specific portions, and wherein each type of capture oligonucleotide in the collection has the same or similar melting temperature but differs substantially in nucleotide sequence, when aligned to another type of capture oligonucleotide in the collection. - View Dependent Claims (9, 10, 11)
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Specification