SHORT CYCLE METHODS FOR SEQUENCING POLYNUCLEOTIDES

• US 20110152114A1
• Filed: 01/18/2011
• Published: 06/23/2011
• Est. Priority Date: 11/12/2003
• Status: Active Grant

• Current Assignee: Fluidigm Corporation
• Original Assignee: Helicos Biosciences Corporation
• Inventors: Buzby, Philip Richard, Harris, Timothy, Lapidus, Stanley
• Granted Patent: US 9,012,144 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• 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 obtaining sequence information from a target nucleic acid, comprising:

a) immobilizing a plurality of different target nucleic acid/primer duplexes at discrete locations on a surface, wherein target nucleic acid portions of said duplexes each comprise the same adaptor sequence, and wherein said primers are hybridized to said adaptor sequence;

b) contacting said plurality of duplexes with a solution comprising an enzyme under conditions wherein the enzyme catalyses template-dependent addition to a terminus of at least one of said primers, resulting in the addition of a label to said primer; and

c) detecting the addition to the terminus of said at least one primer,

wherein said addition is indicative of the presence of a complementary sequence in the target nucleic acid.

28. The method of claim 27, wherein said enzyme is a ligase.

29. The method of claim 27, wherein said enzyme is a polymerase.

30. The method of claim 27, further comprising repeating steps b) and c) one or more times.