Stabilizing a nucleic acid for nucleic acid sequencing

• US 20060252077A1
• Filed: 04/27/2006
• Published: 11/09/2006
• Est. Priority Date: 12/30/2004
• Status: Abandoned Application

• Current Assignee: Fluidigm Corporation
• Original Assignee: Helicos Biosciences Corporation
• Inventors: Buzby, Philip Richard
• Application Number:
  US11/413,381
• Publication Number:
  US 20060252077A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• 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 2533/101 Primer extension
• CPC Class Codes: C12Q 2535/101 Sanger sequencing method, i.e. oligonucleotide sequencing using primer elongation and dideoxynucleotides as chain terminators
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin

Abstract

The invention provides methods for sequencing a nucleic acid comprising stabilizing a primer/target nucleic acid duplex on a substrate. Methods of the invention generally contemplate the use of a dual-anchored primer/target nucleic acid duplex, or a stabilizing molecule in a single molecule sequencing reaction.

Claims

1. A surface for nucleic acid sequencing, said surface comprising a nucleic acid duplex composed of a template and a primer, each of said template and primer being attached to a member of a binding pair, such that each of said members is oriented toward said surface.

2. The surface of claim 1, wherein each duplex is individually optically resolvable.

3. The surface of claim 1, wherein said surface is a polyelectrolyte multilayer.

4. The surface of claim 1, wherein said surface is an epoxide surface.

5. The surface of claim 1, wherein said surface is deposited on a substrate selected from the group consisting of glass and silica.

6. The surface of claim 1, wherein binding partners of said members are covalently attached to said surface.

7. The surface of claim 1, wherein said binding pair is selected from the group consisting of a ligand/receptor pair, an affinity binding pair, an antigen/antibody pair, and a carbohydrate/lectin pair.

8. The surface of claim 7, wherein said pair is selected from the group consisting of biotin/avidin, digoxigenin/anit-digoxigenin, and dintirophenol/anti-dinitrophenol.