LABELLED NUCLEOTIDES

• US 20170002407A1
• Filed: 07/05/2016
• Published: 01/05/2017
• Est. Priority Date: 12/04/2001
• Status: Active Grant

• Current Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Original Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Inventors: BALASUBRAMANIAN, Shankar, BARNES, Colin, LIU, Xiaohai, MILTON, John
• Granted Patent: US 10,480,025 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• 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: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6823 Release of bound markers
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2525/186 incorporating a non-extendable or blocking moiety

Abstract

Nucleosides and nucleotides are disclosed that are linked to detectable labels via a cleavable linker group.

Claims

1-25. -25. (canceled)

26. A nucleotide or nucleoside molecule having a ribose or deoxyribose sugar moiety and a base linked to a detectable label via a cleavable linker, wherein the sugar moiety comprises a protecting group attached via the 3′

oxygen atom, and wherein said protecting group comprises an azido group and can be modified or removed to expose a 3′

OH group.

27. The molecule of claim 26, wherein the base is a purine, or a pyrimidine.

28. The molecule of claim 26, wherein the base is a deazapurine.

29. The molecule of claim 26, which is a deoxyribonucleotide triphosphate.

30. The molecule of claim 26, wherein the detectable label is a fluorophore.

31. The molecule of claim 26, wherein the linker is acid labile, photolabile or contains a disulphide linkage.

32. The molecule of claim 26, wherein the cleavable linker and the protecting group are cleavable under identical conditions.

33. The molecule of claim 26, wherein the protecting group comprises azidomethyl (CH₂N₃).

34. A nucleotide or nucleoside molecule having a ribose or deoxyribose sugar moiety and a base linked to a detectable label via a cleavable linker, wherein the sugar moiety comprises a protecting group attached via the 3′

oxygen atom, and wherein said protecting group comprises cyanoethyl ((CH₂)₂CN) and can be modified or removed to expose a 3′

OH group.

35. The molecule of claim 34, wherein the base is a purine, or a pyrimidine.

36. The molecule of claim 34, wherein the base is a deazapurine.

37. The molecule of claim 34, which is a deoxyribonucleotide triphosphate.

38. The molecule of claim 34, wherein the detectable label is a fluorophore.

39. The molecule of claim 34, wherein the linker is acid labile, photolabile or contains a disulphide linkage.

40. The molecule of claim 34, wherein the cleavable linker and the protecting group are cleavable under identical conditions.

41. A kit comprising a plurality of nucleotides, each having a ribose or deoxyribose sugar moiety and a base linked to a detectable label via a cleavable linker, wherein the sugar moiety comprises a protecting group attached via the 3′

oxygen atom, and said protecting group comprises an azido group and can be modified or removed to expose a 3′

OH group, and wherein the detectable label linked to each nucleotide can be distinguished upon detection from the detectable labels used for structurally different nucleotides.

42. The kit of claim 41, wherein the cleavable linker and the protecting group are cleavable under identical conditions.

43. The kit of claim 41, wherein the protecting group comprises azidomethyl (CH₂N₃).

44. The kit of claim 41, further comprising an enzyme and buffers appropriate for the action of the enzyme.

45. The kit of claim 44, wherein the enzyme is a terminal transferase, a polymerase or a reverse transcriptase.