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US 11,685,946 B2Granted
Complex surface-bound transposome complexes
AssigneeIllumina Cambridge Limited
InventorsAndrew Slatter, Esther Musgrave-Brown, Susan C. Verity, Niall Anthony Gormley
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Abstract
The present disclosure relates to methods, compositions, and kits for generating a library of tagged nucleic acid fragments without using PCR amplification, including methods and compositions for fragmenting and tagging nucleic acids (e.g., DNA) using transposome complexes immobilized on solid support.
Claims11
An annealed transposon/attachment polynucleotide hybrid comprising:
- (a) a first transposon comprising a 3′ transposon end sequence and a 5′ adaptor sequence;
- (b) a second transposon comprising a 5′ transposon end sequence and a 3′ adaptor sequence, wherein
- i. the second transposon 5′ transposon end sequence is complementary to the first transposon 3′ transposon end sequence, forming a double-stranded transposon end sequence to which the transposase is bound,
- ii. the 5′ adaptor sequence of the first transposon and the 3′ adaptor sequence of the second transposon are non-complementary, and
- iii. the first transposon and the second transposon form a Y-shape;
- and
- (c) an attachment polynucleotide comprising:
- i. an attachment adaptor sequence hybridized to either the first transposon 5′ adaptor sequence or the second transposon 3′ adaptor sequence;
- and
- ii. a biotin linker.
7 more elements
A transposome complex comprising:
- (a) a transposase;
- (b) a first transposon comprising a 3′ transposon end sequence and a 5′ adaptor sequence;
- (c) a second transposon comprising a 5′ transposon end sequence and a 3′ adaptor sequence, wherein
- i. the second transposon 5′ transposon end sequence is complementary to the first transposon 3′ transposon end sequence, forming a double-stranded transposon end sequence to which the transposase is bound,
- ii. the 5′ adaptor sequence of the first transposon and the 3′ adaptor sequence of the second transposon are non-complementary, and
- iii. the first transposon and the second transposon form a Y-shape;
- and
- (d) an attachment polynucleotide comprising:
- i. an attachment adaptor sequence hybridized to either the first transposon 5′ adaptor sequence or the second transposon 3′ adaptor sequence and
- ii. a biotin linker.
7 more elements
An immobilized transposome complex comprising:
- a solid support;
- and
- a transposome complex immobilized to the solid support,
- wherein the transposome complex comprises:
- (a) a transposase;
- (b) a first transposon comprising a 3′ transposon end sequence and a 5′ adaptor sequence;
- (c) a second transposon comprising a 5′ transposon end sequence and a 3′ adaptor sequence, wherein
- i. the second transposon 5′ transposon end sequence is complementary to at least a portion of the first transposon 3′ transposon end sequence, forming a double-stranded transposon end sequence to which the transposase is bound,
- ii. the 5′ adaptor sequence of the first transposon and the 3′ adaptor sequence of the second transposon are non-complementary, and
- iii. the first transposon and the second transposon form a Y-shape;
- and
- (d) an attachment polynucleotide comprising:
- i. an attachment adaptor sequence hybridized to either the first transposon 5′ adaptor sequence or the second transposon 3′ adaptor sequence and
- ii. a biotin linker,
- wherein the biotin linker is immobilized to the solid support.
12 more elements
Dependencies are inferred from the claim text.