Patent ID: 11873480
Assignee: ILLUMINA CAMBRIDGE LIMITED
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of preparing a library of barcoded DNA fragments of a target DNA comprising:
a. binding a target DNA to a plurality of transposome complexes, each transposome complex comprising transposons and transposases,
wherein the transposons comprise transferred strands and non-transferred strands, and
wherein at least one of the transposons of the transposome complex comprises an adaptor sequence capable of hybridizing to a complementary capture sequence;
b. fragmenting the target DNA of step a into a plurality of contiguously-linked, transposed DNA fragments and inserting a plurality of transferred strands into at least one strand of each contiguously-linked transposed DNA fragment of the plurality of contiguously-linked transposed fragments, wherein contiguity of DNA fragments of the target DNA is maintained by the transposases;
c. immobilizing the plurality of contiguously-linked, transposed DNA fragments of step b on a plurality of solid supports by hybridizing the adaptor sequence of the at least one of the transposons to a complementary capture sequence, each of the solid supports in the plurality comprising a plurality of immobilized oligonucleotides, each oligonucleotide of the plurality of immobilized oligonucleotides comprising, in sequential order extending from a surface of the solid support:
(i) a first primer binding site attached to the solid support;
(ii) a first barcode sequence; and
(iii) a complementary capture sequence capable of hybridizing to the adaptor sequence; and
d. attaching the first barcode sequence to one or more of the plurality of contiguously-linked, transposed DNA fragments of step c, thereby producing a library of barcoded, linked DNA fragments,
wherein the first barcode sequences of all of the oligonucleotides of the plurality of oligonucleotides immobilized on a given solid support comprise the same nucleic acid sequence,
wherein a nucleic acid sequence of the first barcode sequence of the oligonucleotides of the plurality of oligonucleotides immobilized on a given solid support in the plurality of the solid supports differs from a nucleic acid sequence of all of the first barcode sequences from other solid supports in the plurality of solid supports, and
wherein the steps (a)-(d) are carried out in a single reaction compartment.