# US 2019/0316112 A1 · Methods of capturing a nucleic acid including a target oligonucleotide sequence and uses thereof

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- Number: US 2019/0316112 A1
- Document ID: US20190316112A1
- Status: Published application
- Kind: A1
- Application number: 15/952,045
- Filed: 2018-04-12
- Published: 2019-10-17
- Earliest claimed: 2018-04-12
- Applicants: CellMax, Ltd.
- Inventors: Rui Mei; Julian Lucas; Pratyush Gupta
- Web page: https://patentgrep.com/patent/US20190316112A1

## CPC

- C12N 15/1003: {Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor}
- C12Q 1/6832: Enhancement of hybridisation reaction
- C12Q 1/686: Polymerase chain reaction [PCR]
- C12Q 1/6876: Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C40B 40/08: Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
- 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 are methods of capturing a nucleic acid comprising a target oligonucleotide sequence from a library of nucleic acid that include: contacting a library of nucleic acids comprising a nucleic acid comprising a target oligonucleotide sequence with a probe comprising a sequence that is complementary to the target oligonucleotide sequence, wherein the contacting is performed in a tetramethylammonium chloride (TMAC)-based buffer at a temperature of about 60° C. to about 70° C., and the contacting results in the hybridization of the target oligonucleotide sequence to the sequence that is complementary to the target oligonucleotide sequence, to thereby generate a hybridization product; and isolating the hybridization product from nucleic acids in the library that do not comprise the target oligonucleotide sequence. Also provided are compositions useful for performing these methods.

## Claims

### Claim 1

A composition comprising a liquid, wherein the liquid comprises:
about 0.5 M to about 8.0 M TMAC;
about 10 mM to about 200 mM Tris (pH 8.0);
about 1× to about 5×Denhardt's solution;
about 0.01% to about 0.2% Tween-20;
about 0.5 mM to about 15 mM EDTA; and
about 0.5% to about 25% formamide (v/v).

### Claim 2 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 2.0 M to about 6.0 M TMAC.

### Claim 3 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 5.0 M to about 6.0 M TMAC.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 5.4 M TMAC.

### Claim 5 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 40 mM to about 60 mM Tris (pH 8.0).

### Claim 6 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 100 mM Tris (pH 8.0).

### Claim 7 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 2× to about 3×Denhardt's Solution.

### Claim 8 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 5×Denhardt's Solution.

### Claim 9 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 0.01% to about 0.05% Tween-20.

### Claim 10 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 5 mM to about 15 mM EDTA.

### Claim 11 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 10% to about 25% formamide (v/v).

### Claim 12 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 20% formamide (v/v).

### Claim 13 (depends on 1)

The composition of claim 1, wherein the liquid comprises about 2.7 M TMAC, about 50 mM Tris (pH 8.0), about 2.5×Denhardt's Solution, about 0.010% Tween-20, about 6 mM EDTA, and about 20% formamide.

### Claim 14 (depends on 1)

The composition of claim 1, wherein the composition consists of the liquid.

### Claim 15 (depends on 1)

A kit comprising any one of the compositions of claim 1.