# US 11,299,776 B2 · Methods and devices related to amplifying nucleic acid at a variety of temperatures

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- Number: US 11,299,776 B2
- Document ID: US11299776B2
- Status: Granted
- Kind: B2
- Application number: 16/612,020
- Filed: 2018-05-10
- Published: 2020-08-13
- Granted: 2022-04-12
- Earliest claimed: 2017-05-10
- Assignees: Board of Regents, The University of Texas System
- Applicants: Board of Regents, the University of Texas System
- Inventors: Andrew Ellington; Cheulhee Jung; Sheng Cai; Sanchita Bhadra; John N. Milligan; Daniel Garry; Raghav Shroff
- Web page: https://patentgrep.com/patent/US11299776B2

## CPC

- C12Q 1/6853: using modified primers or templates
- C12Q 1/701: {Specific hybridization probes}
- C12N 9/1252: {DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase}
- C12Q 1/686: Polymerase chain reaction [PCR]
- C12Q 2521/101: DNA polymerase
- C12Q 2525/301: Hairpin oligonucleotides
- C12Q 2527/101: Temperature
- C12Q 2531/101: Linear amplification, i.e. non exponential
- C12Q 2537/1373: Displacement by a nucleic acid
- C12Q 2600/158: Expression markers
- 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

Disclosed are compositions and methods for nucleic acid amplification and detection. Specifically, disclosed herein are compositions and methods that allow for amplification of nucleic acids at a wide variety of temperatures. This includes a polymerase which is thermostable at high temperatures, and a method of amplification that can be conducted at relatively low temperatures.

## Claims

### Claim 1

A non-naturally occurring thermostable polymerase, wherein the thermostable polymerase has at least 97% sequence identity to SEQ ID NO: 1.

### Claim 2 (depends on 1)

The non-naturally occurring thermostable polymerase of claim 1, wherein the polymerase is stored in a storage buffer, or a reaction buffer.

### Claim 3 (depends on 2)

The non-naturally occurring thermostable polymerase of claim 2, wherein the buffer further comprises temperature dependent inhibitor of polymerase activity.

### Claim 4 (depends on 1)

A nucleic acid encoding the non-naturally occurring thermostable polymerase of claim 1.

### Claim 5 (depends on 1)

A kit comprising the non-naturally occurring thermostable polymerase of claim 1.

### Claim 6 (depends on 5)

The kit of claim 5, wherein the kit further comprises at least four distinct primers.

### Claim 7 (depends on 6)

The kit of claim 6, wherein at least one of the primers is phosphorothioated.

### Claim 8 (depends on 5)

The kit of claim 5, wherein the kit further comprises a buffer solution.