Linked ligation
AssigneeNcan Genomics, Inc.
InventorsAndrea Marziali, Joel Pel
Life of this patent
Estimating the term…
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.
Claims21
1-20. (canceled)
A composition, comprising:
- a first nucleic acid strand;
- a second nucleic acid strand, wherein a first segment of the second nucleic acid strand is configured to anneal to a segment of the first nucleic acid strand to define a first duplex region;
- a third nucleic acid strand;
- and
- a fourth nucleic acid strand, wherein a first segment of the fourth nucleic acid strand is configured to anneal to a segment of the third strand to define a second duplex region, wherein a second segment of the fourth nucleic acid strand is configured to anneal to a second segment of the second nucleic acid strand to thereby associate the first duplex region with the second duplex region.
A composition for preparing a sequencing library, the composition comprising:
- first and second oligonucleotides, wherein a 3′ portion of the first oligonucleotide is configured to anneal to a 5′ portion of the second oligonucleotide to form a first adaptor structure comprising a first duplex end portion, and wherein a 5′ portion of the first oligonucleotide and a 3′ portion of the second oligonucleotide are not complementary to each other;
- and
- third and fourth oligonucleotides, wherein a 3′ portion of the third oligonucleotide is configured to anneal to a 5′ portion of the fourth oligonucleotide to form a second adaptor structure comprising a second duplex end portion, and wherein a 5′ portion of the third oligonucleotide and a 3′ portion of the fourth oligonucleotide are not complementary to each other,
- wherein the 3′ portion of the fourth oligonucleotide is configured to anneal to the 3′ portion of the second oligonucleotide.
Dependencies are inferred from the claim text.