Method and apparatus for detecting disorders in genomic substanc

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 18, 436800, 436151, 536 2431, 935 78, 935 76, C12Q 168, C12Q 134, G01N 2518, C07H 1512

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051943727

ABSTRACT:
Disorders of the base sequences in genomic substances such as DNA and RNA are detected by changing the state of aggregation of fine particles by cleavaging using a nuclease. A single-stranded denatured product of the objective genomic substance is added to first and second fine particles each attached to plural pieces of first and second single-stranded nucleic acid probes, respectively. The first and second single-stranded nucleic acid probes are complementary to a first region and a second region, respectively, on the objective genomic substance, which are exclusive of each other and contiguous from each other. Aggregations of the first and second fine particles are formed by double or multiple hybridization reaction of the denatured objective genomic substance added with the first and second single-stranded nucleic acid probes. The aggregations are then digested with a nuclease which cleaves the non-complementary-mismatch-containing portion of the double strand of each hybrid permitting the formation of the aggregations, in the vicinity of a mismatch-localized region, but substantially not the completely complementary portion of the double strand of the hybrid. The size of the aggregations in the digested solution is first measured and then the degree of mismatch between the complementary base sequences of the objective genomic substance and the first or second single-stranded nucleic acid probe is measured using the size of the aggregations.

REFERENCES:
patent: 4711955 (1987-12-01), Word et al.
Brumbaugh et al., DBA Accession No.: 87-12683, Abstr. Pap. Am. Chem. Soc. (193 Meet., ENVR15) 1987.
Lopez-Galindez et al., Proc. Natl. Acad. Sci. USA 85(10), 1988 pp. 3522-3526.
Brumbaugh et al.: Proc. Natl. Acad. Sci., USA 85, 5610-5614, Aug., 1988.

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