WO1996036731A3 - Nucleic acid detection methods - Google Patents

Nucleic acid detection methods Download PDF

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Publication number
WO1996036731A3
WO1996036731A3 PCT/US1996/006527 US9606527W WO9636731A3 WO 1996036731 A3 WO1996036731 A3 WO 1996036731A3 US 9606527 W US9606527 W US 9606527W WO 9636731 A3 WO9636731 A3 WO 9636731A3
Authority
WO
WIPO (PCT)
Prior art keywords
target
nucleic acid
hybridized
nuclease
target sequence
Prior art date
Application number
PCT/US1996/006527
Other languages
French (fr)
Other versions
WO1996036731A2 (en
Inventor
Cassandra L Smith
Ron Yaar
Przemyslaw Szafranski
Charles R Cantor
Original Assignee
Univ Boston
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Univ Boston filed Critical Univ Boston
Priority to AU62486/96A priority Critical patent/AU6248696A/en
Priority to EP96921212A priority patent/EP0827551B1/en
Priority to DE69603723T priority patent/DE69603723T2/en
Publication of WO1996036731A2 publication Critical patent/WO1996036731A2/en
Publication of WO1996036731A3 publication Critical patent/WO1996036731A3/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6823—Release of bound markers

Abstract

The invention relates to methods for rapidly determining the sequence and/or length of a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3'-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguished from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated.
PCT/US1996/006527 1995-05-19 1996-05-20 Nucleic acid detection methods WO1996036731A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU62486/96A AU6248696A (en) 1995-05-19 1996-05-20 Nucleic acid detection methods
EP96921212A EP0827551B1 (en) 1995-05-19 1996-05-20 Nucleic acid detection methods
DE69603723T DE69603723T2 (en) 1995-05-19 1996-05-20 METHOD FOR DETECTING NUCLEIC ACIDS

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/446,102 US5753439A (en) 1995-05-19 1995-05-19 Nucleic acid detection methods
US08/446,102 1995-05-19

Publications (2)

Publication Number Publication Date
WO1996036731A2 WO1996036731A2 (en) 1996-11-21
WO1996036731A3 true WO1996036731A3 (en) 1997-02-06

Family

ID=23771327

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/006527 WO1996036731A2 (en) 1995-05-19 1996-05-20 Nucleic acid detection methods

Country Status (7)

Country Link
US (1) US5753439A (en)
EP (1) EP0827551B1 (en)
AT (1) ATE183244T1 (en)
AU (1) AU6248696A (en)
CA (1) CA2221467A1 (en)
DE (1) DE69603723T2 (en)
WO (1) WO1996036731A2 (en)

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Cited By (2)

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Publication number Priority date Publication date Assignee Title
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US8999266B2 (en) 2000-10-30 2015-04-07 Agena Bioscience, Inc. Method and apparatus for delivery of submicroliter volumes onto a substrate

Also Published As

Publication number Publication date
DE69603723T2 (en) 2000-03-02
ATE183244T1 (en) 1999-08-15
WO1996036731A2 (en) 1996-11-21
AU6248696A (en) 1996-11-29
EP0827551A2 (en) 1998-03-11
US5753439A (en) 1998-05-19
DE69603723D1 (en) 1999-09-16
EP0827551B1 (en) 1999-08-11
CA2221467A1 (en) 1996-11-21

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