Microarray nuclease protection assay
OwnerPaul Kenneth Wolberinventor
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Abstract
A method for microarray analysis of a sample for a target nucleic acid sequence including selecting a microarray comprising a set of oligonucleotide probes, contacting the set of oligonucleotide probes with a sample suspected of containing a target nucleic acid sequence under hybridization conditions to form a set of probe-target hybrids, contacting the set of probe-target hybrids with one or more single-stranded nucleases, removing the target nucleic acids from the remaining set of probe-target hybrids to expose the oligonucleotide probes remaining on the microarray, and labeling the remaining oligonucleotide probes, and analyzing the microarray for oligonucleotide probes hybridized to the target nucleic acid sequences.
Claims21
A method for microarray analysis of a sample for a target nucleic acid sequence, the method comprising:
- selecting a microarray comprising a set of oligonucleotide probes;
- contacting the set of oligonucleotide probes with a sample suspected of containing a target nucleic acid sequence under hybridization conditions to form a set of probe-target hybrids;
- contacting the set of probe-target hybrids with one or more single-stranded nucleases;
- removing the target nucleic acids from the remaining set of probe-target hybrids to expose the oligonucleotide probes remaining on the microarray;
- and
- labeling the remaining oligonucleotide probes;
- and
- analyzing the microarray for oligonucleotide probes hybridized to the target nucleic acid sequences.
A method for microarray analysis of one or more target RNA from a sample, the method comprising
- hybridizing a sample containing one or more target RNAs to a microarray of DNA probes to form a set of target-probe hybrids;
- digesting any unbound and poorly hybridized DNA probes with a single-stranded DNA nuclease;
- removing target RNAs from target-probe hybrids to expose a set of DNA probes on the microarray;
- labeling the DNA probes;
- and
- analyzing the DNA probes remaining on the microarray to gain information about one or more of target RNAs.
A kit for microarray analysis of a sample for a target RNA sequence on a microarray comprising a set of DNA oligonucleotide probes, the kit comprising:
- one or more single-stranded DNA nucleases;
- one or more RNases;
- and
- one or more labeling moieties for labeling DNA oligonucleotide probes;
- and
- instructions for use of the nucleases, RNases and labeling moieties on a microarray for analysis of a sample for a target RNA sequence.
Dependencies are inferred from the claim text.