Sequencing method using magnifying tags

• US 9,458,493 B2
• Filed: 03/02/2011
• Issued: 10/04/2016
• Est. Priority Date: 12/23/1998
• Status: Expired due to Fees

• Current Assignee: Empire IP LLC
• Original Assignee: Empire IP LLC
• Inventors: Lexow, Preben
• Primary Examiner(s): Whisenant, Ethan C
• Application Number:
  US13/039,197
• Publication Number:
  US 20150247181A1
• Time in Patent Office: 2,043 Days
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/313 Type II endonucleases, i.e. cutting outside recognition site
• CPC Class Codes: C12Q 2521/501 Ligase
• CPC Class Codes: C12Q 2525/155 incorporating/generating a new priming site
• CPC Class Codes: C12Q 2525/185 incorporating bases where the precise position of the bases in the nucleic acid string is important
• CPC Class Codes: C12Q 2525/191 incorporating an adaptor

Abstract

The present invention provides a method of sequencing all or part of a target nucleic acid molecule by determining the sequence of a portion of said nucleic acid molecule in addition to information regarding the position of said portion and in particular provides a new method of sequencing involving the magnification of one or more bases of said bases to aid identification.

Claims

1. A method for magnifying a signal associated with at least one base in all or part of a target nucleic acid molecule, comprising:

a) converting at least a portion of a target nucleic acid molecule to a single strand suitable for binding an adapter molecule;

b) binding to at least a portion of the single strand an adapter molecule comprising one or more magnifying tags corresponding to one or more bases of the single strand,

c) ligating the adapter molecule to the single strand;

d) binding to at least a further portion of the single strand a further adapter molecule comprising one or more further magnifying tags corresponding to one or more further bases of the single strand; and

e) ligating the adapter molecule to the further adapter molecule to form a complementary strand.

2. The method for magnifying a signal associated with at least one base in all or part of a target nucleic acid molecule according to claim 1, further comprising:

f) repeating steps b) to e) to form a double strand.

3. The method for magnifying a signal associated with at least one base in all or part of a target nucleic acid molecule according to claim 2, further comprising:

g) denaturing the double strand; and

h) analyzing the magnifying tags in the complementary strand to determine a sequence of bases in the target nucleic acid molecule.