Homogeneous polynucleotide displacement assay method kit and reagent complex
A method is described for determining the presence of a predetermined target nucleotide sequence in the DNA of a biological sample which comprises the steps: (a) providing a reagent complex of (i) a probe polynucleotide which is capable of base pair binding via hydrogen bonds of purine/pyrimidine bases to the DNA target nucleotide sequence, and (ii) an RNA signal strand polynucleotide which is bound by base pair binding via hydrogen bonds of purine/pyrimidine base pairs to the probe polynucleotide in a region of the probe polynucleotide at least partially coextensive with the region in which the probe polynucleotide is capable of binding to the DNA target nucleotide sequence; (b) contacting the reagent complex with a sample under conditions in which the DNA target nucleotide sequence, if present, binds to the probe polynucleotide and displaces the RNA signal strand polynucleotide from the reagent complex; (c) without separation, digesting the displaced RNA signal strand polynucleotide selectively with respect to RNA signal strand polynucleotide remaining in reagent complex; and (d) detecting the presence of the digestion products of digesting the displaced RNA signal strand polynucleotide.
- Assignee:
- Allied Corp., Morris Township, NJ
- Patent Number(s):
- US 4795701
- OSTI ID:
- 6465893
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
BINDING ENERGY
BIOCHEMISTRY
CHEMICAL BONDS
CHEMISTRY
COMPLEXES
DIGESTION
DNA SEQUENCING
ELEMENTS
ENERGY
HYDROGEN
INTRACELLULAR DIGESTION
MOLECULAR BIOLOGY
NONMETALS
NUCLEIC ACIDS
NUCLEOTIDES
ORGANIC COMPOUNDS
REAGENTS
RNA
STRAND BREAKS
STRUCTURAL CHEMICAL ANALYSIS