WO2001068919A3 - Mutation detection using denaturing gradients - Google Patents

Mutation detection using denaturing gradients Download PDF

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Publication number
WO2001068919A3
WO2001068919A3 PCT/US2001/007906 US0107906W WO0168919A3 WO 2001068919 A3 WO2001068919 A3 WO 2001068919A3 US 0107906 W US0107906 W US 0107906W WO 0168919 A3 WO0168919 A3 WO 0168919A3
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WO
WIPO (PCT)
Prior art keywords
polynucleotide
wild type
target
mutant
probe
Prior art date
Application number
PCT/US2001/007906
Other languages
French (fr)
Other versions
WO2001068919A2 (en
Inventor
Saeedullah Mohammad Raees
Brian Alfred Perry
Original Assignee
Ana Gen Technologies Inc
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 Ana Gen Technologies Inc filed Critical Ana Gen Technologies Inc
Priority to AU2001245650A priority Critical patent/AU2001245650A1/en
Publication of WO2001068919A2 publication Critical patent/WO2001068919A2/en
Publication of WO2001068919A3 publication Critical patent/WO2001068919A3/en

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    • 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

Abstract

Methods for detecting single nucleotide polymorphisms in a sample by applying an energy gradient to a mixture of probe-hybridized target wild type and mutant polynucleotides are provided. A nucleotide probe, having a nucleotide sequence complementary to at least a portion of the wild type or mutant polynucleotide(s), is exposed to a sample containing target wild type polynucleotide(s) and target mutant polynucleotide(s) under hybridizing conditions. Target wild type polynucleotide(s) form homoduplexes with the wild type probe, while target mutated polynucleotide(s) form heteroduplexes with the wild type probe. Target mutant polynucleotide(s) form homoduplexes with the mutant probes, while target wild type polynucleotide(s) form heteroduplexes with the mutant probe. An en ergy gradient is applied to the duplex-containing sample to induce selective denaturation of the duplexes. The gradient may be a thermal or chemical one and is preferably temporally linear. The hetero- and homoduplexes deanneal at different times so that the detection of more than one type of polynucleotide(s) is indicative of a single nucleotide polymorphism. Capillary electropheresis, gel electropheresis, high performance liquid chromatography, or microfluidics are typically used for identification of the separated strands.
PCT/US2001/007906 2000-03-10 2001-03-12 Mutation detection using denaturing gradients WO2001068919A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001245650A AU2001245650A1 (en) 2000-03-10 2001-03-12 Mutation detection using denaturing gradients

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US18853400P 2000-03-10 2000-03-10
US60/188,534 2000-03-10

Publications (2)

Publication Number Publication Date
WO2001068919A2 WO2001068919A2 (en) 2001-09-20
WO2001068919A3 true WO2001068919A3 (en) 2002-12-05

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2001/007906 WO2001068919A2 (en) 2000-03-10 2001-03-12 Mutation detection using denaturing gradients

Country Status (3)

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US (2) US6596487B2 (en)
AU (1) AU2001245650A1 (en)
WO (1) WO2001068919A2 (en)

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Also Published As

Publication number Publication date
US20020042060A1 (en) 2002-04-11
US6596487B2 (en) 2003-07-22
US20030175798A1 (en) 2003-09-18
WO2001068919A2 (en) 2001-09-20
AU2001245650A1 (en) 2001-09-24

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