IP Library › Granted Patent US 12,509,723
Granted Patent B2
US 12,509,723 · App. 18/489,381 · Granted Dec 30, 2025

Methods of sequencing with linked fragments

Inventors: Milenko Despotovic (Richmond, CA); Joel Pel (Vancouver, CA); Andrea Marziali (North Vancouver, CA)
Assignee: NCAN GENOMICS, INC.
C12Q1/6869
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Quick Facts
Patent No.
US 12,509,723
App. No.
18/489,381
Granted
Dec 30, 2025
Kind
B2
Abstract

The invention generally relates to sequencing library preparation methods. In certain embodiments, two template nucleic acids are joined together by a linking molecule, such as a PEG derivative. The linked template nucleic acids is amplified, creating linked amplicons.

Claims (16)

1 . A method of sequencing a nucleic acid, the method comprising:

seeding a single amplification reaction with a first nucleic acid molecule and a second nucleic acid molecule, wherein the second nucleic acid molecule has a sequence that is identical except for any errors introduced during sample preparation to a sequence of the first nucleic acid molecule or a reverse complement thereof;

performing the amplification reaction on the first nucleic acid molecule and the second nucleic acid molecule to generate amplicons; and

conducting a sequencing reaction with the amplicons to determine a consensus sequence of the first nucleic acid molecule and the second nucleic acid molecule.

2 . The method of claim 1 , wherein the second nucleic acid molecule is a first strand copy that was synthesized by extending a polymerase to copy a complementary strand to the first nucleic acid molecule.

3 . The method of claim 2 , wherein the amplicons are attached to a solid support.

4 . The method of claim 1 , wherein the first nucleic acid molecule and the second nucleic acid molecule represent both senses of a fragment of DNA.

5 . The method of claim 1 , further comprising, prior to the sequencing step, making the first nucleic acid molecule and the second nucleic acid molecule by providing linked primers comprising at least a first primer linked to a second primer; annealing the first primer to a template of interest; extending the first primer; annealing the second primer to the template; and extending the second primer.

6 . The method of claim 5 , wherein the first primer and the second primer bind to universal binding sites.

7 . The method of claim 1 , wherein the amplification reaction includes emulsion PCR.

8 . The method of claim 1 , wherein the amplification reaction comprises bridge amplification.

9 . The method of claim 8 , wherein the sequencing reaction generates sequence data and the method includes detecting, from the sequencing data, polymerase error that occurred during the amplification reaction.

10 . The method of claim 1 , wherein the first nucleic acid molecule and the second nucleic acid molecule are made by extending linked primers to copy a template of interest.

11 . The method of claim 10 , wherein the first nucleic acid molecule and the second nucleic acid molecule are linked by a linker.

12 . The method of claim 10 , wherein the linked primers are linked by a linker that includes polyethylene glycol (PEG), modified PEG, an inverted base, or a modified base.

13 . The method of claim 10 , wherein the linked primers include adaptors compatible with a nucleic acid sequencing system.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 31, 2023
From: DESPOTOVIC, MILENKO; PEL, JOEL; MARZIALI, ANDREA
To: BOREAL GENOMICS INC.
Reel/Frame 065405/0556 →
CHANGE OF NAME Recorded Oct 31, 2023
From: BOREAL GENOMICS INC.
To: NCAN GENOMICS, INC.
Reel/Frame 065405/0671 →
Continuity (5)
Continuation 17060853 · Oct 1, 2020
Continuation 16008702 · Jun 14, 2018
Continuation 14930227 · Nov 2, 2015
Provisional Application 62074991 · Nov 4, 2014
Related Publication 20240043921A1 · Feb 8, 2024
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