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US 11,306,348 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.
Claims30
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 the 5′ transposon end sequence is complementary to the 3′ transposon end sequence;
- wherein the 5′ adaptor sequence is non-complementary to the 3′ adaptor sequence;
- and
- wherein the first transposon and the second transposon form a Y-shape;
- and
- d. an attachment polynucleotide comprising:
- i. an attachment adaptor sequence, wherein the attachment adaptor sequence is complementary to and hybridized to one of the two adaptor sequences, and
- ii. a binding element, optionally wherein the binding element comprises biotin, optionally wherein the binding element in a 3′ biotin linker;
- and
- wherein the transposome complex comprises at least one uracil.
10 more elements
Dependencies are inferred from the claim text.