Modular nucleotide compositions and uses therefor

• US 8,846,881 B2
• Filed: 07/19/2012
• Issued: 09/30/2014
• Est. Priority Date: 11/19/2008
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Korlach, Jonas, Wegener, Jeffrey
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US13/552,952
• Publication Number:
  US 20130022968A1
• Time in Patent Office: 803 Days
• Field of Search: 536/4.1, 536/26.6, 435/5, 435/6.1, 435/7.1, 435/7.2, 435/91.1
• US Class Current: 536/4.1
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• 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: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites

Abstract

Nucleic acid compositions, methods of making and using such compositions that comprise modular functional groups that can be configured to provide desired functionality to different nucleotide types, through a swappable and preferably non-covalent linkage component. Such compositions are useful in a variety of applications including nucleic acid analyses.

Claims

1. A composition comprising a reaction component coupled to a fluorescent label through a non-covalent linkage component configured to provide a separation between the fluorescent label and the reaction component, wherein the reaction component comprises a nucleoside polyphosphate, and further wherein the fluorescent label causes photo-induced damage to an enzyme interacting with the reaction component under excitation illumination in the absence of the separation provided by the non-covalent linkage component, which shields the enzyme from the fluorescent label during interaction between the enzyme and the reaction component.

2. The composition of claim 1, wherein the linkage component comprises a sacrificial component that is impacted by the photo-induced damage, thereby shielding the enzyme.

3. The composition of claim 1, wherein the linkage component comprises a triplet state quencher.

4. The composition of claim 1, wherein the linkage component is a multivalent linkage group.

5. The composition of claim 4, wherein the multivalent linkage group couples multiple functional groups to the reaction component, wherein the fluorescent label is among the multiple functional groups.

6. The composition of claim 4, wherein the multivalent linkage group is selected from avidin, streptavidin, and neutravidin.

7. The composition of claim 4, wherein the fluorescent label comprises multiple fluorophores, and the multivalent linkage group is coupled to the multiple fluorophores.

8. The composition of claim 4, wherein the multivalent linkage group is coupled to both the fluorescent label and to a photo-induced damage mitigating agent.

9. The composition of claim 1, wherein the fluorescent label is a FRET label.

10. The composition of claim 1, wherein the linkage component is coupled to a phosphate group of the nucleoside polyphosphate.

11. The composition of claim 10, wherein the linkage component is coupled to a phosphate group of the nucleoside polyphosphate by a non-covalent linkage.

12. The composition of claim 1, wherein the enzyme is a polymerase enzyme.

13. A method of shielding an enzyme from photo-induced damage within a reaction mixture, the method comprising dispensing the composition of claim 1 into the reaction mixture.

14. The composition of claim 1, further dispensed into a reaction mixture comprising the enzyme.