LABELED REACTANTS AND THEIR USES

• US 20110311964A1
• Filed: 05/25/2011
• Published: 12/22/2011
• Est. Priority Date: 03/13/2008
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Wegener, Jeffrey, Korlach, Jonas
• Granted Patent: US 8,354,252 B2
• Time in Patent Office: Days
• US Class Current: 435/6.1
• 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: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2563/107 fluorescence

Abstract

Labeled reactant compositions, and particularly labeled nucleic acid reaction compositions, that include structural components that maintain potentially damaging labeling components sufficiently distal from the reactant portion of the molecule such that damaging effects of the label group on other reaction components, such as enzymes, are reduced, minimized and/or eliminated.

Claims

1-29. -29. (canceled)

30. A method of monitoring a nucleic acid synthesis reaction, comprising:

contacting a polymerase/template/primer complex with a fluorescently labeled nucleotide or nucleotide analog having a nucleotide or nucleotide analog component, a fluorescent label component, and a linker component joining the nucleotide or nucleotide analog component to the label component, wherein the linker component maintains the label component at a functional distance away from the nucleotide or nucleotide analog component of at least 2 nm; and

detecting a characteristic signal from the fluorescent dye indicative of incorporation of the nucleotide or nucleotide analog into a primer extension reaction.

32. The method of claim 30, wherein the functional distance is at least 5 nm.

33. The method of claim 30, wherein the functional distance is at least 10 nm.

34. The method of claim 30, wherein the linker component comprises groups selected from conjugated alkynes, conjugated alkenes, aryl alkynes, aryl amides, polyethylene glycol, oligopeptide, nucleic acids, PNA, and LNA.

35. The method of claim 30, wherein the linker component comprises an oligopeptide.

36. The method of claim 35, wherein the oligopeptide comprises oligoproline.

37. The method of claim 30, wherein the contacting step is carried out under excitation illumination for the label component.

38. The method of claim 30, wherein the detecting step is carried out in real time during a template mediated primer extension reaction.

39. The method of claim 30, wherein the polymerase primer template complex is provided immobilized upon a solid support.

40. The method of claim 39, wherein the solid support comprises a zero mode waveguide, and the complex is immobilized within the zero mode waveguide.

31. (canceled)