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-001/68
C12M-001/00
C12M-001/34
C12M-003/00
출원번호
US-0252169
(2008-10-15)
등록번호
US-8492085
(2013-07-23)
발명자
/ 주소
Barany, Francis
Zirvi, Monib
Gerry, Norman P.
Favis, Reyna
Kliman, Richard
출원인 / 주소
Cornell Research Foundation, Inc.
대리인 / 주소
LeClairRyan, a Professional Corporation
인용정보
피인용 횟수 :
4인용 특허 :
62
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 first step of the method involves providing a first set of a plurality of tetramers of four nucleotides linked together, where (1) each tetramer within the first set differs from all other tetramers in the first set by at least two nucleotide bases, (2) no two tetramers within the first set are complementary to one another, (3) no tetramers within the first set are palindromic or dinucleotide repeats, and (4) no tetramer within the first set has one or less or three or more G or C nucleotides. Groups of 2 to 4 of the tetramers from the first set are linked together to form a collection of multimer units. From the collection of multimer units, all multimer units formed from the same tetramer and all multimer units having a melting temperature in .degree. C. of less than 4 times the number of tetramers forming a multimer unit are removed to form a modified collection of multimer units. The modified collection of multimer units is arranged in a list in order of melting temperature. The order of the modified collection of multimer units is randomized in 2.degree. C. increments of melting temperature.
대표청구항▼
1. An oligonucleotide array comprising: a support anda collection of double multimer unit oligonucleotides at different positions on the support so that complementary oligonucleotides to be immobilized on the support can be captured at the different positions, wherein the complementary oligonucleoti
1. An oligonucleotide array comprising: a support anda collection of double multimer unit oligonucleotides at different positions on the support so that complementary oligonucleotides to be immobilized on the support can be captured at the different positions, wherein the complementary oligonucleotides will hybridize, within a temperature range of greater than 24° C., to members of the collection of double multimer unit oligonucleotides, the double multimer unit oligonucleotides are formed from sets of four to eight tetramers where (1) each tetramer within a set differs from all other tetramers in the set by at least two nucleotide bases, (2) no two tetramers within a set are complementary to one another, and (3) no tetramers within a set are palindromic or dinucleotide repeats, and the collection of double multimer unit oligonucleotides has had the following oligonucleotides removed from it: (1) oligonucleotides having a melting temperature in ° C. less than 11 times the number of tetramers and more than 15 times the number of tetramers, (2) double multimer units with the same 3 tetramers linked together, and (3) double multimer units with the same 4 tetramers linked together with or without interruption. 2. An oligonucleotide array according to claim 1, wherein the collection of double multimer units is shown in FIG. 26. 3. An oligonucleotide array according to claim 1, wherein the collection of double multimer units is shown in FIG. 27. 4. An oligonucleotide array according to claim 1, wherein double multimer unit oligonucleotides having a melting temperature in ° C. less than 12.5 times the number of tetramers and more than 14 times the number of tetramers are removed from the collection of double multimer unit oligonucleotides. 5. An oligonucleotide array according to claim 1, wherein the double multimer units are 24 mers and the melting temperature of the double multimer units is 75-84° C. 6. An oligonucleotide array according to claim 1, wherein the set of tetramers is shown in Table 6 and complements thereof. 7. An oligonucleotide array according to claim 1, wherein the set of tetramers are one base circular permutations of the tetramers shown in Table 6 and complements thereof. 8. A oligonucleotide array according to claim 1, wherein the set of tetramers are two base circular permutations of the tetramers shown in Table 6 and complements thereof. 9. An oligonucleotide array according to claim 1, wherein the set of tetramers are three base circular permutations of the tetramers shown in Table 6 and complements thereof. 10. A kit for identifying one or more of a plurality of sequences differing by single-base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences comprising: a ligase;a plurality oligonucleotide probe sets, each characterized by (a) a first oligonucleotide probe, having a target sequence-specific portion and an addressable array-specific portion, and (b) a second oligonucleotide probe, having a target sequence-specific portion and detectable reporter label, wherein the oligonucleotide probes in a particular set are suitable for ligation together when hybrided adjacent to one another on a respective target nucleotide sequence, but have a mismatch which interferes with such ligation when hybridized to any other nucleotide sequence, present in the sample; anda support with different capture oligonucleotides immobilized at different positions, wherein the capture oligonucleotides have nucleotide sequences complementary to the addressable array-specific portions and are formed from a collection of double multimer unit oligonucleotides, wherein oligonucleotides with addressable array-specific portions will hybridize, within a temperature range of greater than 4 times the number of tetramers in the double multimer unit, to members of the capture oligonuncleotides, the double multimer unit oligonucleotides are formed from sets of four to eight tetramers where (1) each tetramer within a set differs from all other tetramers in the set by at least two nucleotide bases, (2) no two tetramers within a set are complementary to one another, and (3) no tetramers within a set are palindromic or dinucleotide repeats, and the collection of double multimer unit oligonucleotides has had the following oligonucleotides removed from it: (1) oligonucleotides having a melting temperature in ° C. of less than 11 times the number of tetramers and more than 15 times the number of tetramers, (2) double multimer units with the same 3 tetramers linked together, and (3) double multimer units with the same 4 tetramers linked together with or without interruption, wherein the capture oligonucleotides have nucleotide sequences complementary to the addressable array-specific portions. 11. A kit according to claim 10, wherein the mismatch of oligonucleotide probe sets to nucleotide sequences other than their respective target nucleotide sequences is at a base at a ligation junction at which the oligonucleotide probe of each set ligate together when hybridized to their respective target nucleotide sequences. 12. A kit according to claim 10, wherein the mismatch is on the oligonucleotide probe of the oligonucleotide probe sets which have 3′ ends at the ligation junction. 13. A kit according to claim 10, wherein the mismatch of oligonucleotide probe sets to nucleotide sequences other than their respective target nucleotide sequence is at a base adjacent to a ligation junction at which the oligonucleotide probes of each set ligate together when hybridized to their respective target nucleotide sequences. 14. A kit according to claim 13, wherein the mismatch is on the oligonucleotide probe of the oligonucleotide probe sets which have 3′ ends at the ligation junction. 15. A kit according to claim 10, wherein the ligase is selected from the group consisting of Thermus aquaticus ligase, Thermus thermophilus ligase, E. coli ligase, T4 ligase, and Pyrococcus ligase. 16. A kit according to claim 10 further comprising: amplification primers suitable for preliminary amplification of the target nucleotide sequences anda polymerase. 17. A kit according to claim 10, wherein the support includes different capture oligonucleotides immobilized at different particular sites with different capture oligonucleotides being complementary to different addressable array-specific portions, whereby different oligonucleotide probe sets are hybridized and detected at different sites on the support. 18. A kit according to claim 10, wherein the support includes identical capture oligonucleotides immobilized on the support with the capture oligonucleotides complementary to all the addressable array-specific portions and the labels attached to the oligonucleotide probe sets being different, whereby the oligonucleotide probe sets are detected and distinguished by the different labels. 19. A kit according to claim 10, wherein the oligonucleotide probe sets and the capture oligonucleotides are configured so that the oligonucleotide probe sets hybridize, respectively, to the target nucleotide sequences at temperatures which are less than that at which the capture oligonucleotides hybridize to the addressable array-specific portions of the oligonucleotide probes sets. 20. A kit according to claim 10, wherein the collection of double multimer units is shown in FIG. 26. 21. A kit according to claim 10, wherein the collection of double multimer units is shown in FIG. 27. 22. A kit according to claim 10, wherein double multimer unit oligonucleotides having a melting temperature in ° C. of less than 12.5 times the number of tetramers and more than 14 times the number of tetramers are removed from the collection of double multimer unit oligonucleotides. 23. A kit according to claim 10, wherein the double multimer units are 24 mers and the melting point of the double multimer units is 75-84° C. 24. A kit according to claim 10, wherein the set of tetramers is shown in Table 1 or complements thereof. 25. A kit according to claim 10, wherein the set of tetramers are one base circular permutations of the tetramers shown in Table 1 and complements thereof. 26. A kit according to claim 10, wherein the set of tetramers are two base circular permutations of the tetramers shown in Table 1 and complements thereof. 27. A kit according to claim 10, wherein the set of tetramers are three base circular permutations of the tetramers shown in Table 1 and complements thereof.
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