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)
C07H-021/04
C12N-015/11
C12Q-001/68
B01J-019/00
B82Y-030/00
C40B-040/06
C40B-060/14
출원번호
US-0101067
(2013-12-09)
등록번호
US-9234241
(2016-01-12)
발명자
/ 주소
Barany, Francis
Barany, George
Hammer, Robert P.
Kempe, Maria
Blok, Herman
Zirvi, Monib
출원인 / 주소
Cornell Research Foundation, Inc.
대리인 / 주소
LeClairRyan, a Professional Corporation
인용정보
피인용 횟수 :
0인용 특허 :
118
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.
대표청구항▼
1. A method comprising: providing a collection of oligonucleotides wherein each type of oligonucleotide is greater than 16 nucleotides and differs in nucleotide sequence, when aligned to another type of oligonucleotide in the collection, by at least 25%;providing a sample comprising a plurality of c
1. A method comprising: providing a collection of oligonucleotides wherein each type of oligonucleotide is greater than 16 nucleotides and differs in nucleotide sequence, when aligned to another type of oligonucleotide in the collection, by at least 25%;providing a sample comprising a plurality of composite oligonucleotides, each composite oligonucleotide comprising (i) a zip-code portion, and (ii) one or more further nucleotide sequences;contacting the sample comprising the plurality of composite oligonucleotides with the collection of oligonucleotides under conditions effective to hybridize the zip-code portion of each composite oligonucleotide to its complementary oligonucleotide in the collection; anddetecting the one or more of the plurality of composite oligonucleotides hybridized to their complementary oligonucleotides in the collection. 2. The method according to claim 1, wherein each oligonucleotide in the collection comprises 20-25 nucleotides. 3. The method of according to claim 1, wherein the collection of oligonucleotides is coupled to a solid support. 4. The method according to claim 3, wherein said detecting is carried out on the solid support. 5. The method according to claim 1, wherein the zip-code portions of the composite oligonucleotides hybridize to their complementary oligonucleotides in the collection under stringent hybridization conditions. 6. The method according to claim 1, wherein the collection contains at least 25 types of oligonucleotides. 7. The method according to claim 1, wherein the collection contains at least 1000 types of oligonucleotides. 8. The method according to claim 1, wherein the one or more further nucleotide sequences of the composite oligonucleotide comprises an amplification product, or complement thereof, of one or more cellular target nucleic acid sequences. 9. A method comprising: providing a solid support having an array of positions, wherein composite oligonucleotides are coupled to the solid support at the array of positions, each of said composite oligonucleotides characterized by having a target-specific portion and a zip code portion, wherein the zip code portion of each of the composite oligonucleotides comprises a nucleotide sequence of greater than sixteen nucleotides that differs for each different target-specific portion, wherein the nucleotide sequence of one zip code portion differs from the nucleotide sequence of another zip code portion, when aligned to each other, by at least 25% of the nucleotides; anddetecting one or more labels coupled to at least one of the composite oligonucleotides, thereby facilitating identification of the composite oligonucleotide and its zip code portion. 10. The method of claim 9, wherein said detecting is carried out in an imager. 11. The method of claim 10, wherein the imager is a fluorescence imager and the detectable label is a fluorescent label suitable for detection by the fluorescence imager. 12. The method of claim 9, wherein the composite oligonucleotides are coupled to beads. 13. The method of claim 9, wherein different composite oligonucleotides are coupled to the solid support at different array positions. 14. The method of claim 9, wherein the solid support having an array of positions comprises a high-density array of composite oligonucleotides immobilized thereto. 15. The method of claim 9, 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. 16. The method of claim 9, wherein the solid support is in a form selected from the group consisting of slides, discs, membranes, films, and composites thereof. 17. The method of claim 9, wherein the target nucleotide sequence in the sample comprises an amplification product, or complement thereof, of one or more cellular target nucleic acid sequences. 18. The method of claim 9, wherein the zip-code portion of the composite oligonucleotide comprises a nucleotide sequence of 24 nucleotides or less. 19. An apparatus comprising: a solid support having an array of positions;composite oligonucleotides coupled to the solid support at the array of positions, each of said composite oligonucleotides characterized by having a target-specific portion and a zip code portion, wherein the zip code portion of each of the composite oligonucleotides comprises a nucleotide sequence of greater than sixteen nucleotides, wherein the nucleotide sequence of one zip code portion differs from the nucleotide sequence of another zip code portion, when aligned to each other, by at least 25% of the nucleotides; andone or more detectable labels coupled to at least one of the composite oligonucleotides. 20. The apparatus of claim 19 further comprising: an imager for detecting or quantifying at least one of the detectable labels at at least one of the array positions on the solid support. 21. The apparatus of claim 19, wherein the composite oligonucleotides are coupled to beads. 22. The apparatus of claim 19, wherein the solid support having an array of positions comprises a high-density array of composite oligonucleotides immobilized thereto. 23. The apparatus of claim 19, wherein the target nucleotide sequence in the sample comprises an amplification product, or complement thereof, of one or more cellular target nucleic acid sequences.
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