Phospholink nucleotides for sequencing applications

• US 8,367,813 B2
• Filed: 11/17/2009
• Issued: 02/05/2013
• Est. Priority Date: 11/17/2008
• 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
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US12/620,293
• Publication Number:
  US 20100167299A1
• Time in Patent Office: 1,176 Days
• Field of Search: 536/4.1, 536/23.1, 536/25.3, 536/26.6, 435/6, 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 1/6874 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 2563/155 Particles of a defined size, e.g. nanoparticles
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence

Abstract

The present invention provides labeled phospholink nucleotides that can be used in place of naturally occurring nucleotide triphosphates or other analogs in template directed nucleic acid synthesis reactions and other nucleic acid reactions and various analyzes based thereon, including DNA sequencing, single base identification, hybridization assays, and others.

Claims

1. A composition, comprising a labeled phospholink nucleotide, wherein said labeled phospholink nucleotide comprises a structure that has formula:

2. The composition of claim 1, wherein said linker moiety is a member selected from substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocycloalkyl.

3. The composition of claim 1, wherein said linker moiety comprises poly(ethylene glycol).

4. The composition of claim 1, wherein said bead is a latex bead.

5. The composition of claim 1, wherein said bead is a fluorescent bead.

8. A kit comprising at least four differently labeled nucleotide analogs each comprising said structure of said composition of claim 1, a polymerase enzyme source, and instructions for using said composition with said polymerase enzyme in template directed nucleic acid synthesis.

6. A composition, comprising a labeled phospholink nucleotide, wherein said labeled phospholink nucleotide comprises a structure that has formula:

7. The composition of claim 6, wherein said polymeric material is a member selected from polyethylene glycol and alginate.

9. The composition of claim 6, wherein said linker moiety is a member selected from substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocycloalkyl.

10. The composition of claim 6, wherein said linker moiety comprises poly(ethylene glycol).

11. The composition of claim 6, wherein said bead is a latex bead.

12. The composition of claim 6, wherein said bead is a fluorescent bead.

13. A kit comprising at least four differently labeled nucleotide analogs each comprising said structure of said composition of claim 6, a polymerase enzyme source, and instructions for using said composition with said polymerase enzyme in template directed nucleic acid synthesis.