# US 10,246,736 B2 · Methods and apparatuses for nucleic acid shearing by sonication

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- Number: US 10,246,736 B2
- Document ID: US10246736B2
- Status: Granted
- Kind: B2
- Application number: 14/816,857
- Filed: 2015-08-03
- Published: 2015-11-26
- Granted: 2019-04-02
- Earliest claimed: 2008-02-15
- Assignees: Life Technologies Corporation
- Applicants: Life Technologies Corporation
- Inventors: Vladimir I. Bashkirov; Umberto Ulmanella; Robert G. Eason; Bradford J. Taft
- Web page: https://patentgrep.com/patent/US10246736B2

## CPC

- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- C07H 1/06: Separation; Purification
- C07H 21/00: Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C12N 15/1093: {General methods of preparing gene libraries, not provided for in other subgroups}
- 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
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Methods and kits for preparing nucleic acid fragments from a sample of purified nucleic acid are provided. Alternatively, chromatin or other long polymers can be sheared with similar methods and kits.

## Claims

### Claim 1

A method for preparing a gene fragment library, the method comprising:
a) providing a suspension comprising a sample of purified genomic nucleic acid and monodisperse particles, wherein the suspension is an aqueous solution without nonaqueous solvent:
b) sonicating the suspension, thereby generating nucleic acid fragments:
c) collecting the nucleic acid fragments; and
d) preparing a gene fragment library by ligating the nucleic acid fragments to nucleic acid adaptors.

### Claim 2 (depends on 1)

The method of claim 1, wherein the collecting step does not include chemical extraction of the nucleic acid fragments.

### Claim 3 (depends on 1)

The method of claim 1, wherein the sample does not include cell lysates.

### Claim 4 (depends on 1)

The method of claim 1, wherein the particles have a size of about 0.1 micrometers to about 100 micrometers.

### Claim 5 (depends on 1)

The method of claim 1, wherein the suspension contains about 0.1% to about 20% of the particles.

### Claim 6 (depends on 1)

The method of claim 1, wherein the particles comprise at least one of glass, borosilicate glass, silica, soda lime glass, carbamide, metal, metal oxide, ceramic, and metal-coated ceramic.

### Claim 7 (depends on 1)

The method of claim 1, wherein the particles are non-magnetic or paramagnetic.

### Claim 8 (depends on 1)

The method of claim 1, wherein the sonicating step is performed by focusing acoustic sonication.

### Claim 9 (depends on 1)

The method of claim 1, wherein the time for the sonicating step is less than 5 minutes to generate about 60 to about 90 base pair nucleic acid fragments from the purified genomic nucleic acid.

### Claim 10 (depends on 1)

The method of claim 1, further comprising sequencing the nucleic acid fragments.

### Claim 11 (depends on 1)

The method of claim 1, further comprising hybridizing the nucleic acid fragments to target nucleic acids located on a microarray.