Method of designing addressable array suitable for detection of nucleic acid sequence differences using ligase detection reaction
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/6837
C12Q-001/6827
C12Q-001/6874
C12Q-001/6886
출원번호
US-0644273
(2017-07-07)
등록번호
US-10131938
(2018-11-20)
발명자
/ 주소
Barany, Francis
Zirvi, Monib
Gerry, Norman P.
Favis, Reyna
Kliman, Richard
출원인 / 주소
Cornell Research Foundation, Inc.
대리인 / 주소
LeClairRyan PLLC
인용정보
피인용 횟수 :
0인용 특허 :
68
초록▼
The present invention is directed to a method of designing a plurality of capture oligonucleotide probes for use on a support to which complementary oligonucleotide probes will hybridize with little mismatch, where the plural capture oligonucleotide probes have melting temperatures within a narrow r
The present invention is directed to a method of designing a plurality of capture oligonucleotide probes for use on a support to which complementary oligonucleotide probes will hybridize with little mismatch, where the plural capture oligonucleotide probes have melting temperatures within a narrow range. The present invention further relates to an oligonucleotide array comprising of a support with the plurality of oligonucleotide probes immobilized on the support, a method of using the support to detect single-base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences, and a kit for such detection, which includes the support on which the oligonucleotides have been immobilized.
대표청구항▼
1. An apparatus comprising: a solid support having an array of positions; a collection of composite or encoded oligonucleotides coupled to the solid support at the array of positions, each of said composite or encoded oligonucleotides characterized by having a target-specific portion and an address,
1. An apparatus comprising: a solid support having an array of positions; a collection of composite or encoded oligonucleotides coupled to the solid support at the array of positions, each of said composite or encoded oligonucleotides characterized by having a target-specific portion and an address, wherein the address comprises two multimer unit sequences coupled to the solid support, wherein each multimer unit sequence (1) is comprised of three consecutive subunits linked together, each subunit independently selected from the group consisting of a trimer, tetramer, pentamer, or hexamer sequence, (2) differs from other multimer unit sequences by at least two nucleotide bases in at least one of the subunits when aligned, and (3) has a Tm value greater than 24° C., wherein the collection of oligonucleotides has at least 64 different types of oligonucleotides, and wherein the collection of composite or encoded oligonucleotides does not contain: (1) composite or encoded oligonucleotides having an address with a melting temperature in ° C. less than 11 times the number of subunits, (2) composite or encoded oligonucleotides having an address comprised of two identical multimer unit sequences linked together and (3) two types of composite or encoded oligonucleotides with an address having the same four subunit sequences linked together in the same order without interruption; and one or more detectable labels coupled to at least one of the composite or encoded oligonucleotides, thereby facilitating detection of the composite or encoded oligonucleotide and its address. 2. The apparatus of claim 1 further comprising: an imager for detecting or quantifying at least one of the detectable labels at least one of the array positions on the solid support. 3. The apparatus of claim 2, wherein the composite or encoded oligonucleotides are coupled to a group of spheres. 4. The apparatus of claim 2, wherein different composite or encoded oligonucleotides are coupled to the solid support at different array positions. 5. The apparatus of claim 2, wherein the solid support having an array of positions comprises an array of composite or encoded oligonucleotides immobilized thereto. 6. The apparatus of claim 2, wherein the solid support is made from a material selected from the group consisting of plastic, ceramic, metal, gel, glass, silicon, and composites thereof. 7. The apparatus of claim 2, wherein the solid support is in a form selected from the group consisting of slides, discs, membranes, and films. 8. The apparatus of claim 2, wherein the address of the composite or encoded oligonucleotide comprises a nucleotide sequence that is complementary to a secondary oligonucleotide probe, and the target specific sequence comprises a nucleotide sequence that is the same as, or complementary to, a target nucleotide sequence in a sample. 9. The apparatus of claim 8, wherein the target nucleotide sequence in the sample comprises an amplification product, or complement thereof, of one or more target nucleic acid sequences. 10. The apparatus of claim 2, wherein the imager is a fluorescence imager and the detectable label is a fluorescent label suitable for detection by the fluorescence imager. 11. The apparatus of claim 2, wherein the one or more detectable labels is indicative of the target-specific sequence included within the composite or encoded oligonucleotide to which it is coupled. 12. The apparatus of claim 2, wherein the one or more detectable labels is indicative of a nucleic acid sequence in a sample that is the same as, or complementary to, the target-specific sequence included within the composite or encoded oligonucleotide to which it is coupled. 13. The apparatus of claim 2, wherein the one or more detectable labels is coupled to the address of the composite or encoded oligonucleotide in a target specific manner by an enzyme. 14. The apparatus of claim 2, wherein detection of one or more detectable labels coupled to the address of the composite or encoded oligonucleotides correlates to the presence of one or more target nucleic acid sequences in the sample that is the same as, or complementary to the target-specific portion of the composite or encoded oligonucleotide. 15. The apparatus of claim 2, wherein detection, identification, and quantification of the one or more detectable labels correlates to the presence, identity, or quantity of one or more target nucleic acid sequences in a sample. 16. The apparatus of claim 3, wherein the group of spheres has greater than 4,000 composite or encoded oligonucleotides. 17. A device comprising: a solid support having an array of positions each suitable for attachment of one or more of a collection of oligonucleotides; an array of oligonucleotides on the solid support at the array of positions, wherein the one or more of a collection of oligonucleotides on the solid support comprises two multimer unit sequences coupled to the solid support, wherein each multimer unit sequence is comprised of three consecutive subunits linked together, each subunit independently selected from the group consisting of a trimer, tetramer, pentamer, or hexamer sequence, wherein each multimer unit sequence has a Tm value of greater than 24° C., wherein each of the two multimer unit sequences differs from all other multimer unit sequences by at least two nucleotide bases in at least one subunit when aligned, wherein the collection of oligonucleotides has at least 64 different types of oligonucleotides, and wherein the collection of oligonucleotides does not contain: (1) oligonucleotides having a sequence with a melting temperature in ° C. less than 11 times the number of subunits, (2) oligonucleotides having a sequence comprised of two identical multimer unit sequences linked together and (3) two types of oligonucleotides with a sequence having the same four subunit sequences linked together in the same order without interruption, wherein one or more of the oligonucleotides of the collection is hybridized to a nucleic acid molecule comprising a complementary nucleotide sequence; and wherein one or more target nucleic acid molecules is coupled to one or more of the oligonucleotides of the collection. 18. A device according to claim 17, wherein identifying the oligonucleotides at the array of positions facilitates identifying the one or more target nucleic acid molecules. 19. The device according to claim 17 wherein the array has greater than 4,000 oligonucleotides. 20. The device according to claim 17, wherein different oligonucleotides are attached at different array positions on the solid support. 21. The device according to claim 17, wherein oligonucleotides having the same sequences are attached at different positions on the support. 22. The device according to claim 17, wherein the solid support is in a form selected from the group consisting of particles, strands, precipitates, gels, sheets, tubing, spheres, containers, capillaries, pads, slices, films, plates, slides, discs, membranes, or any combination or composite thereof. 23. The device according to claim 17, wherein a linker couples each of the one or more oligonucleotides at the array of positions to the solid support. 24. An instrument comprising: a solid support comprising at least 64 types of oligonucleotides coupled to the solid support at an array of positions, wherein one or more oligonucleotides coupled to the solid support at a position on the array comprises an address that comprises two multimer unit sequences coupled to the solid support, wherein each multimer unit sequence (1) is comprised of three consecutive subunits linked together, each subunit independently selected from the group consisting of a trimer, tetramer, pentamer, or hexamer sequence, (2) differs from the other multimer unit sequences by at least two nucleotide bases in at least one of the subunits when aligned, and (3) has a Tm value greater than 24° C., and wherein the oligonucleotides on the array do not contain: (1) oligonucleotides having an address with a melting temperature in ° C. less than 11 times the number of subunits, (2) oligonucleotides having an address comprised of two identical multimer unit sequences linked together and (3) two types of oligonucleotides with an address having the same four subunit sequences linked together in the same order without interruption; one or more nucleic acid molecules hybridized to complementary portions of the one or more oligonucleotides on said solid support; and an imager configured to detect the hybridized nucleic acid molecules. 25. The instrument of claim 24, wherein the solid support comprises an array of greater than 4,000 oligonucleotides immobilized thereto. 26. The instrument of claim 24, wherein the one or more oligonucleotides immobilized on or coupled to the solid support at a position on the array comprises a target-specific portion and an address. 27. The instrument of claim 24, wherein the one or more nucleic acid molecules comprise amplification products, or complements thereof, of one or more target nucleic acid molecules. 28. The apparatus of claim 1, wherein the imager is a fluorescence imager and the nucleic acid molecules hybridized to the one or more oligonucleotides on the solid support each comprise a fluorescent label suitable for detection by the fluorescence imager.
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