Single-stranded labelled oligonucleotides of preselected sequence

• US 5,541,313 A
• Filed: 11/09/1994
• Issued: 07/30/1996
• Est. Priority Date: 02/22/1983
• Status: Expired due to Term

• Current Assignee: Molecular Biosystems, Inc (Alliance Pharmaceutical Corporation)
• Original Assignee: Molecular Biosystems, Inc (Alliance Pharmaceutical Corporation)
• Inventors: Ruth, Jerry L.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): CRANE, LAWRENCE E
• Application Number:
  US08/336,500
• Time in Patent Office: 629 Days
• Field of Search: 536/24.3, 536/24.31, 536/24.32, 435/6
• US Class Current: 536/24.3
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6813 Hybridisation assays

Abstract

Substantially pure single-stranded oligonucleotides having a preselected sequence of not more than about 200 nucleotides, at least one of which is at a preselected position in the sequence and includes a base with a covalently attached linker arm containing or capable of binding at least one reporter group or solid support. A process for the chemical synthesis of the substantially pure single-stranded oligonucleotide and modified nucleosides useful in such synthesis are provided.

Claims

1. A composition consisting essentially of single-stranded oligonucleotides, wherein the sequence of each of said oligonucleotides is substantially identical to the sequence of each of the other oligonucleotides in said composition,

wherein each of said oligonucleotides has a sequence of not more than approximately 200 nucleotides,

wherein at least one of the nucleotide monomers from which each of said oligonucleotides is prepared has a linker arm attached to the base of said nucleotide, said nucleotide being substantially identically located in each of said oligonucleotides, and

wherein said linker arm is further attached to a reporter group.

2. A composition according to claim 1 wherein said linker arm is attached at a sterically tolerant site.

3. A composition according to claim 1 wherein said base is a pyrimidine and said linker arm is attached to the C-5 position thereof.

4. A composition according to claim 1 wherein said base is a purine and said linker arm is attached to the C-8 position thereof.

5. A composition according to claim 1 wherein said identical sequence is between about 10 to about 40 nucleotides in length.

6. A composition according to claim 1 wherein said reporter group is a functional colorimetric group.

7. A composition according to claim 1 wherein said reporter group is a functional fluorescent group.

8. A composition according to claim 1 wherein said reporter group is a functional luminescent group.

9. A composition according to claim 1 wherein said reporter group is a functional radioactive group.

10. A composition according to claim 1 wherein said reporter group is a functional ligand recognition group.

11. A composition according to claim 1 wherein said reporter group is blocked.

12. A composition of single-stranded oligonucleotides wherein the sequence of each of the oligonucleotides in said composition is substantially identical to the sequence of each of the other oligonucleotides in said composition,

wherein each of said oligonucleotides has a sequence of not more than approximately 200 nucleotides,

wherein at least one of the nucleotides of each of said oligonucleotides has a linker arm attached to the base of said nucleotide, said nucleotide being substantially identically located in each of said oligonucleotides, and

wherein said linker arm is further attached to a reporter group.

13. The composition of claim 12 wherein said linker arm is attached at a sterically tolerant site.

14. The composition of claim 12 wherein said base is a pyrimidine and said linker arm is attached to the C-5 position thereof.

15. The composition of claim 12 wherein said base is a purine and said linker arm is attached to the C-8 position thereof.

16. The composition of claim 12 wherein said identical sequence is between about 10 to about 40 nucleotides in length.

17. A composition according to claim 12 wherein said reporter group is a functional colorimetric group.

18. A composition according to claim 12 wherein said reporter group is a functional fluorescent group.

19. A composition according to claim 12 wherein said reporter group is a functional luminescent group.

20. A composition according to claim 12 wherein said reporter group is a functional radioactive group.

21. A composition according to claim 12 wherein said reporter group is a functional ligand recognition group.

22. A composition according to claim 12 wherein said reporter group is blocked.

23. A composition consisting essentially of single-stranded oligonucleotides,

wherein the sequence of each of said oligonucleotides is substantially identical to the sequence of each of the other oligonucleotides in said composition,

wherein each of said oligonucleotides has a sequence of not more than approximately 200 nucleotides,

wherein at least one of the nucleotide monomers from which each of said oligonucleotides is prepared has a linker arm attached to the base of said nucleotide, said nucleotide being substantially identically located in each of said oligonucleotides, and

wherein said linker arm is further attached to a solid support.

24. A composition according to claim 23 wherein said linker arm is attached at a sterically tolerant site.

25. A composition according to claim 23 wherein said base is a pyrimidine and said linker arm is attached to the C-5 position thereof.

26. A composition according to claim 23 wherein said base is a purine and said linker arm is attached to the C-8 position thereof.

27. A composition according to claim 23 wherein said identical sequence is between about 10 to about 40 nucleotides in length.

28. A composition of single-stranded oligonucleotides wherein the sequence of each of the oligonucleotides in said composition is substantially identical to the sequence of each of the other oligonucleotides in said composition,

wherein each of said oligonucleotides has a sequence of not more than approximately 200 nucleotides,

wherein at least one of the nucleotides of each said oligonucleotides has a linker arm attached to the base of said nucleotide, said nucleotide being substantially identically located in each of said oligonucleotides, and

wherein said linker arm is further attached to a solid surface.

29. A composition according to claim 28 wherein said linker arm is attached at a sterically tolerant site.

30. A composition according to claim 28 wherein said base is a pyrimidine and said linker arm is attached to the C-5 position thereof.

31. A composition according to claim 28 wherein said base is a purine and said linker arm is attached to the C-8 position thereof.

32. A composition according to claim 28 wherein said identical sequence is between about 10 to about 40 nucleotides in length.