SHORT CYCLE METHODS FOR SEQUENCING POLYNUCLEOTIDES

• US 20110151449A1
• Filed: 01/18/2011
• Published: 06/23/2011
• Est. Priority Date: 11/12/2003
• Status: Abandoned Application

• Current Assignee: Fluidigm Corporation
• Original Assignee: Helicos Biosciences Corporation
• Inventors: Buzby, Phillip Richard, Harris, Timothy, Lapidus, Stanley
• Application Number:
  US13/008,468
• Publication Number:
  US 20110151449A1
• Time in Patent Office: Days
• US Class Current: 435/6.1
• 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 2527/113 Time

Abstract

The invention provides methods for sequencing a polynucleotide comprising stopping an extension cycle in a sequence by synthesis reaction before the reaction has run to near or full completion.

Claims

1-26. -26. (canceled)

27. A method for sequencing a nucleic acid template, the method comprising the steps of:

(a) extending a primer hybridized to a nucleic acid template in a primer/template duplex by exposing said primer to (i) a polymerase capable of catalyzing nucleotide addition to said primer, and (ii) one or more extendible nucleotides each comprising a label, under conditions wherein said polymerase extends said primer by incorporation of a nucleotide;

(b) identifying said nucleotide incorporated into said primer;

(c) repeating said extending and said identifying steps to determine the nucleotide sequence of the nucleic acid molecule.

28. The method of claim 27, wherein said one or more extendible nucleotides comprises a single type of nucleotide.

29. The method of claim 27, wherein said detectable label comprises an optically detectable label.

30. The method of claim 29, wherein said optically detectable label comprises a fluorescent label.

31. The method of claim 29, wherein said optically detectable label comprises a chemiluminescent label.

32. The method of claim 27, wherein said identifying comprising detecting label on said nucleotide incorporated into said primer.

33. The method of claim 27, further comprising a step wherein unbound nucleotides are removed after said extending and prior to said identifying.