Polynucleotide determination with selectable cleavage sites
First Claim
1. A method for detecting the presence of an oligonucleotide sequence of interest in a nucleitic acid analyte present in a nucleic acid sample, said method comprising:
- combining under hybridizing conditions said nucleic acid sample with a polynucleotide reagent, wherein one of said sample or a component of said reagent is bound to a support and hybridization of said analyte and said polynucleotide regent results in a label being bound to said support through a selectable cleavage site;
substantially freeing said support of label bound to said support other than through said selectable cleavage site;
cleaving said cleavage site; and
detecting the presence of freed label.
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Accused Products
Abstract
Novel methods for assaying a nucleic acid analyte are provided, which employ polynucleotides having oligonucleotide sequences substantially homologous to a sequence of interest in the analyte, where the presence or absence of hybridization at a predetermined stringency provides for the release of a label from a support. Particularly, various techniques are employed for binding a label to a support, whereupon cleavage of either a single or double strand, a label may be released from a support, where the release of the label can be detected as indicative of the presence of a particular oligonucleotide sequence in a sample. The method finds use in diagnosis of disease, genetic monitoring, and analysis of nucleic acid mixtures.
397 Citations
12 Claims
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1. A method for detecting the presence of an oligonucleotide sequence of interest in a nucleitic acid analyte present in a nucleic acid sample, said method comprising:
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combining under hybridizing conditions said nucleic acid sample with a polynucleotide reagent, wherein one of said sample or a component of said reagent is bound to a support and hybridization of said analyte and said polynucleotide regent results in a label being bound to said support through a selectable cleavage site; substantially freeing said support of label bound to said support other than through said selectable cleavage site; cleaving said cleavage site; and detecting the presence of freed label. - View Dependent Claims (2, 5, 7, 9, 11)
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3. A method for detecting the presence of an oligonucleotide sequence of interest in a nucleitic acid analyte present in a nucleic acid sample, said method comprising:
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combining under hybridizing conditions said nucleic acid sample with a polynucleotide reagent, where one of said sample or a component of said reagent is bound to a support and hybridization of said analyte and said polynucleotide regent results in a label being bound to said support through a selectable cleavage site; separating said support having bound polynucleotide reagent and nucleic acid analyte from said aqueous medium; washing said support with a medium of different hybridizing stringency from said aqueous medium to remove label bound to said support other than through said selectable cleavage site; cleaving said cleavage site; and detecting the presence of feed label. - View Dependent Claims (4, 6, 8, 10, 12)
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Specification