Linked ligation
AssigneeBoreal Genomics, Inc.
InventorsAndrea Marziali, Joel Pel
Life of this patent
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Abstract
The invention generally relates to capturing, amplifying, and sequencing nucleic acids. In certain embodiments, copies of the sense and antisense strands of a duplex template nucleic acid are captured using linked capture probes and multiple binding and extension steps to improve specificity over traditional single binding target capture techniques. Methods of seeding sequencing clusters with sense and antisense strands of a target nucleic acid are also disclosed including identifying the strands using sense-specific barcodes and confirming base calls using two sense-specific sequencing reads. Linked adapters may be used to increase adapter ligation selectively or efficiency and yield.
Claims25
A method of selectively ligating adapters to a target nucleic acid, the method comprising:
- providing a first linked ligation adapter comprising a probe complimentary to a first portion of a target nucleic acid, the probe linked to a first adapter comprising a first universal priming site;
- exposing a sample comprising the target nucleic acid to the first linked ligation adapter;
- ligating the target nucleic acid to the first linked adapter;
- and
- amplifying the ligated target nucleic acid by PCR using a first universal primer complimentary to the first universal priming site.
A method of ligating double stranded adapters to a template DNA molecule, the method comprising:
- providing a linked adapter comprising a first double stranded adapter linked by a linker to a second double stranded adapter;
- exposing the linked adapter to a template DNA molecule;
- ligating the first adapter to one end of the template DNA molecule;
- and
- ligating the second adapter to another end of the template DNA molecule.
Dependencies are inferred from the claim text.