# US 10,865,410 B2 · Next-generation sequencing libraries

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- Number: US 10,865,410 B2
- Document ID: US10865410B2
- Status: Granted
- Kind: B2
- Application number: 16/023,574
- Filed: 2018-06-29
- Published: 2018-11-22
- Granted: 2020-12-15
- Earliest claimed: 2013-08-19
- Assignees: Abbott Molecular Inc.
- Applicants: Abbott Molecular Inc.
- Inventors: Dae Hyun Kim
- Web page: https://patentgrep.com/patent/US10865410B2

## CPC

- C12N 15/1086: {Preparation or screening of expression libraries, e.g. reporter assays}
- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- C40B 40/06: Libraries containing nucleotides or polynucleotides, or derivatives thereof
- C40B 50/06: Biochemical methods, e.g. using enzymes or whole viable microorganisms
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C40B: COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES (in silico combinatorial libraries of nucleic acids, proteins or peptides G16B35/00; in silico combinatorial chemistry G16C20/60)

## Abstract

Provided herein is technology relating to next-generation sequencing and particularly, but not exclusively, to methods and compositions for preparing a next-generation sequencing library comprising short overlapping DNA fragments and using the library to sequence one or more target nucleic acids.

## Claims

### Claim 1

A system for sequencing a target nucleic acid, the system comprising:
a) a nucleic acid ladder fragment library wherein said nucleic acid ladder fragment library comprises a plurality of nucleic acid fragments terminated by a 3′-O-propargyl nucleotide; and
b) a sequencing apparatus.

### Claim 2 (depends on 1)

The system of claim 1 further comprising a copper-based click chemistry catalyst reagent.

### Claim 3 (depends on 1)

The system of claim 1 further comprising an adaptor oligonucleotide.

### Claim 4 (depends on 1)

The system of claim 1 further comprising an adaptor oligonucleotide, wherein the 3′-O-propargyl nucleotide comprises a first reactive group and the adaptor oligonucleotide comprises a second reactive group that is capable of being linked to the first reactive group by click chemistry.

### Claim 5 (depends on 1)

The system of claim 1 wherein said nucleic acid ladder fragment ladder library comprises a plurality of nucleic acids having 3′ ends that differ by less than 20 nucleotides.

### Claim 6 (depends on 1)

The system of claim 1 further comprising software for assembling short overlapping nucleotide sequences into a consensus sequence.

### Claim 7 (depends on 6)

The system of claim 6 wherein:
i) each short overlapping nucleotide sequence comprises less than 100 bases;
ii) the short overlapping nucleotide sequences are tiled over a target nucleic acid comprising at least 100 bases; and
iii) the short overlapping nucleotide sequences are offset from one another by 1-20 bases.