# US 10,501,780 B2 · Compositions for in situ nucleic acid analysis

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,501,780 B2
- Document ID: US10501780B2
- Status: Granted
- Kind: B2
- Application number: 15/041,648
- Filed: 2016-02-11
- Published: 2016-08-11
- Granted: 2019-12-10
- Earliest claimed: 2007-05-18
- Assignees: Applied Biosystems, LLC
- Applicants: Applied Biosystems, LLC
- Inventors: Richard A. Fekete; Annalee Nguyen
- Web page: https://patentgrep.com/patent/US10501780B2

## CPC

- C12Q 1/6841: In situ hybridisation
- C12N 15/1003: {Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor}
- C12Q 1/6844: Nucleic acid amplification reactions
- C12Q 1/686: Polymerase chain reaction [PCR]
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Sample preparation processes for in situ RNA or DNA analysis, methods and compositions therefor are provided. Processes provided herein allow DNA or RNA analysis to be carried out in the same tube or on an aliquot of the prepared sample without centrifugation or extraction. The preparation process can be carried out at room temperature in as little as seven minutes and is amenable to high throughput processing using manual or robotic platforms.

## Claims

### Claim 1

A composition comprising: a polypeptide having deoxyribonuclease activity and a surfactant that substantially lacks fluorescence between 300 nm and 750 nm when in use for in situ analysis of nucleic acid, wherein the composition is substantially free of a cation chelator.

### Claim 2 (depends on 1)

The composition of claim 1 further comprising a ribonuclease inhibitor.

### Claim 3 (depends on 1)

The composition of claim 1 further comprising a reverse transcriptase.

### Claim 4 (depends on 1)

The composition of claim 1 further comprising a polymerase.

### Claim 5 (depends on 1)

The composition of claim 1 wherein the surfactant comprises octylphenol ethoxylate having an average of 7.5 ethoxylate groups.

### Claim 6 (depends on 1)

The composition of claim 1 wherein the surfactant comprises octylphenol ethoxylate having an average of 9.5 ethoxylate groups.

### Claim 7 (depends on 1)

The composition of claim 1 wherein the surfactant comprises dodecyl alcohol polyoxyethylene ether having an average of 9.5 ethoxylate groups.

### Claim 8 (depends on 1)

The composition of claim 1 wherein the surfactant comprises octylphenolpoly(ethyleneglycolether).

### Claim 9 (depends on 1)

The composition of claim 1 comprising less than 0.1 mM cation chelator.

### Claim 10 (depends on 1)

The composition of claim 1 comprising less than 0.2 mM cation chelator.

### Claim 11 (depends on 1)

The composition of claim 1 comprising less than 0.5 mM cation chelator.

### Claim 12 (depends on 1)

The composition of claim 1 comprising less than 1 mM cation chelator.

### Claim 13

A composition comprising: a polypeptide having deoxyribonuclease activity and a surfactant that substantially lacks fluorescence between 300 nm and 750 nm when in use for in situ analysis of nucleic acid, wherein the composition is substantially free of a cation chelator, wherein the surfactant comprises octylphenol ethoxylate having an average of 7.5 ethoxylate groups, and wherein the composition comprises less than 1 mM cation chelator.

### Claim 14 (depends on 13)

The composition of claim 13 comprising less than 0.5 mM cation chelator.

### Claim 15 (depends on 13)

The composition of claim 13 comprising less than 0.2 mM cation chelator.

### Claim 16 (depends on 13)

The composition of claim 13 comprising less than 0.1 mM cation chelator.

### Claim 17

A composition comprising: a polypeptide having deoxyribonuclease activity and a surfactant that substantially lacks fluorescence between 300 nm and 750 nm when in use for in situ analysis of nucleic acid, wherein the composition is substantially free of a cation chelator, wherein the surfactant comprises octylphenol ethoxylate having an average of 9.5 ethoxylate groups, and wherein the composition comprises less than 1 mM cation chelator.

### Claim 18 (depends on 17)

The composition of claim 17 comprising less than 0.5 mM cation chelator.

### Claim 19 (depends on 17)

The composition of claim 17 comprising less than 0.2 mM cation chelator.

### Claim 20 (depends on 17)

The composition of claim 17 comprising less than 0.1 mM cation chelator.