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US 5,853,990 AGranted
Real time homogeneous nucleotide assay
AssigneeEdward E. Winger
InventorsEdward E. Winger, Donald J. Kessler, David E. Hargrove
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Abstract
A method for performing real time, homogeneous assay of a target nucleic acid comprising annealing a labeled, ribo-oligonucleotide probe to the target DNA sequence and degrading a portion of the probe with RNase H to release labeled fragments. Sequential measurements of the released fragments permits kinetic characterization of the presence of the target sequence. Preferably, the assay is integrated into a polymerase chain reaction so that target amplification can be detected in real time.
Claims7
A method for the detection of a target DNA sequence in a sample comprising the steps of:
- a) contacting and annealing a probe comprising a ribo-oligonucleotide attached to a first label with a sample containing a target single-stranded DNA sequence having a region complementary to the probe;
- b) cleaving at least one of the ribonucleotides of the annealed probe with a ribo-nucleic acid nuclease capable of hydrolyzing ribonucleotides in a double stranded RNA:DNA duplex to release labeled ribo-oligonucleotide fragments;
- and
- c) generating a signal from the released labeled ribo-oligonucleotide fragments that kinetically characterizes the target DNA sequence;
- wherein steps a)-c) are performed in a single reaction mixture.
2 more elements
Dependencies are inferred from the claim text.