Detection of nucleic acid sequence differences using the ligase detection reaction with addressable arrays
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C07H-021/02
C07H-021/00
G01N-033/543
출원번호
US-0963920
(2001-09-26)
발명자
/ 주소
Barany,Francis
Barany,George
Hammer,Robert P.
Kempe,Maria
Blok,Herman
Zirvi,Monib
출원인 / 주소
Cornell Research Foundation, Inc.
Board of Supervisors of Louisiana State University and Agricultural and Mechanical College
Regents of the University of Minnesota
대리인 / 주소
Nixon Peabody LLP
인용정보
피인용 횟수 :
45인용 특허 :
57
초록▼
The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detectio
The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detection phase. The ligation phase utilizes a ligation detection reaction between one oligonucleotide probe, which has a target sequence-specific portion and an addressable array-specific portion, and a second oligonucleotide probe, having a target sequence-specific portion and a detectable label. After the ligation phase, the capture phase is carried out by hybridizing the ligated oligonucleotide probes to a solid support with an array of immobilized capture oligonucleotides at least some of which are complementary to the addressable array-specific portion. Following completion of the capture phase, a detection phase is carried out to detect the labels of ligated oligonucleotide probes hybridized to the solid support. The ligation phase can be preceded by an amplification process. The present invention also relates to a kit for practicing this method, a method of forming arrays on solid supports, and the supports themselves.
대표청구항▼
What is claimed: 1. A device comprising: a solid support having an array of positions each suitable for attachment of an oligonucleotide probe; a linker suitable for coupling an oligonucleotide probe to the solid support and attached to the solid support at each of the array positions; and an array
What is claimed: 1. A device comprising: a solid support having an array of positions each suitable for attachment of an oligonucleotide probe; a linker suitable for coupling an oligonucleotide probe to the solid support and attached to the solid support at each of the array positions; and an array of capture oligonucleotide probes on the solid support at the array positions, said capture oligonucleotide probes each having greater than sixteen nucleotides and able to bind to complementary nucleic acids at uniform hybridization conditions, wherein each capture oligonucleotide probe of the array differs in sequence from its adjacent capture oligonucleotide probe, when aligned to each other by at least 25% of the nucleotides. 2. The device according to claim 1, wherein different capture oligonucleotide probes are attached at different array positions on the solid support to detect different nucleic acids. 3. The device according to claim 1, wherein the solid support is made from a material selected from the group consisting of plastic, ceramic, metal, resin, gel, glass, silicon, and composites thereof. 4. The device according to claim 1, wherein the solid support is in a form selected from the group consisting of slides, discs, membranes, films, and composites thereof. 5. The device according to claim 1, wherein the solid support has an array of positions with capture oligonucleotide probes attached to all of the array of positions. 6. The device according to claim 5, wherein the solid support has wells, raised regions, or etched trenches. 7. The device according to claim 6, wherein the solid support is in a microtiter plate. 8. The device according to claim 1, wherein the linker comprises a silane on a surface of the solid support. 9. The device according to claim 1, wherein the solid support is functionalized with olefin, amino, hydroxyl, silanol, aldehyde, keto, halo, acyl halide, or carboxyl groups. 10. The device according to claim 9, wherein the solid support is functionalized with an amino group by reaction with an amine compound selected from the group consisting of 3-aminopropyl triethoxysilane, 3-aminopropylmethyldiethoxysilane, 3-aminopropyl dimethylethoxysilane, 3-aminopropyl trimethoxysilane, N-(2-aminoethyl)-3-aminopropylmethyl dimethoxysilane, N-(2-aminoethyl-3-aminopropyl) trimethoxysilane, aminophenyl trimethoxysilane, 4-aminobutyldimethyl methoxysilane, 4-aminobutyl triethoxysilane, aminoethylaminomethylphenethyl trimethoxysilane, and mixtures thereof. 11. The device according to claim 9, wherein the solid support is functionalized with an olefin-containing silane. 12. The device according to claim 11, wherein the olefin-containing silane is selected from the group consisting of 3-(trimethoxysilyl)propyl methacrylate, N-[3-(trimethoxysilyl)propyl]-N'-(4-vinylbenzyl)ethylenediamine, triethoxyvinylsilane, triethylvinylsilane, vinyltrichlorosilane, vinyltrimethoxysilane, vinyltrimethylsilane, and mixtures thereof. 13. The device according to claim 11, wherein the silanized support is polymerized with an olefin containing monomer. 14. The device according to claim 13, wherein the olefin-containing monomer contains a functional group. 15. The device according to claim 14, wherein the olefin-containing monomer is selected from the group consisting of acrylic acid, methacrylic acid, vinylacetic acid, 4-vinylbenzoic acid, itaconic acid, allyl amine, allylethylamine, 4-aminostyrene, 2-aminoethyl methacrylate, acryloyl chloride, methacryloyl chloride, chlorostyrene, dichlorostyrene, 4-hydroxystyrene, hydroxymethylstyrene, vinylbenzyl alcohol, allyl alcohol, 2-hydroxyethyl methacrylate, poly(ethylene glycol) methacrylate, and mixtures thereof. 16. The device according to claim 13, wherein the support is polymerized with a monomer selected from the group consisting of acrylic acid, acrylamide, methacrylic acid, vinylacetic acid, 4-vinylbenzoic acid, itaconic acid, allyl amine, allylethylamine, 4-aminostyrene, 2-aminoethyl methacrylate, acryloyl chloride, methacryloyl chloride, chlorostyrene, dichlorostyrene, 4-hydroxystyrene, hydroxymethyl styrene, vinylbenzyl alcohol, allyl alcohol, 2-hydroxyethyl methacrylate, poly(ethylene glycol) methacrylate, and mixtures thereof, together with a monomer selected from the group consisting of acrylic acid, methacrylic acid, vinylacetic acid, 4-vinylbenzoic acid, itaconic acid, allyl amine, allylethylamine, 4-aminostyrene, 2-aminoethyl methacrylate, acryloyl chloride, methacryloyl chloride, chlorostyrene, dichlorostyrene, 4-hydroxystyrene, hydroxymethyl styrene, vinylbenzyl alcohol, allyl alcohol, 2-hydroxyethyl methacrylate, poly(ethylene glycol) methacrylate, methyl acrylate, methyl methacrylate, ethyl acrylate, ethyl methacrylate, styrene, 1-vinylimidazole, 2-vinylpyridine, 4-vinylpyridine, divinylbenzene, ethylene glycol dimethacrylate, N,N'-methylenediacrylamide, N,N'-phenylenediacrylamide, 3,5-bis(acryloylamido) benzoic acid, pentaerythritol triacrylate, trimethylolpropane trimethacrylate, pentaerytrithol tetraacrylate, trimethylolpropane ethoxylate (14/3 EO/OH) triacrylate, trimethylolpropane ethoxylate (7/3 EO/OH) triacrylate, trimethylolpropane propoxylate (1 PO/OH) triacrylate, trimethylolpropane propoxylate (2 PO/OH) triacrylate, and mixtures thereof. 17. The device according to claim 1, wherein the linker or support is non-hydrolyzable. 18. The device according to claim 1, wherein the array is reusable for repeatedly hybridizing oligonucleotides to the array of capture oligonucleotides probes on the solid support. 19. The device according to claim 1, wherein each of the capture oligonucleotide probes on the solid support has a different nucleotide sequence.
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Fodor Stephen P. A. (Palo Alto CA) Pirrung Michael C. (Durham NC) Read J. Leighton (Palo Alto CA) Stryer Lubert (Stanford CA), Array of oligonucleotides on a solid substrate.
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Ekins Roger P. (Department of Molecular Endocrinology University College and Middlesex School of Medicine Mortimer Street London W1N 8AA GBX) Chu Frederick W. (Department of Molecular Endocrinology U, Binding assay employing labelled reagent.
Fraiser Melinda S. (Durham NC) Walker George T. (Chapel Hill NC) Schram James L. (Knightdale NC), Decontamination of nucleic acid amplification reactions using uracil-N-glycosylase (UDG).
Barany, Francis; Gerry, Norman P.; Witowski, Nancy E.; Day, Joseph; Hammer, Robert P.; Barany, George, Detection of nucleic acid sequence differences using the ligase detection reaction with addressable arrays.
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