# US 10,370,702 B2 · Methods of making oligonucleotide probes

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- Number: US 10,370,702 B2
- Document ID: US10370702B2
- Status: Granted
- Kind: B2
- Application number: 15/273,041
- Filed: 2016-09-22
- Published: 2017-01-05
- Granted: 2019-08-06
- Earliest claimed: 2012-10-18
- Assignees: President and Fellows of Harvard College
- Applicants: President and Fellows of Harvard College
- Inventors: Chao-ting Wu; Brian Beliveau
- Web page: https://patentgrep.com/patent/US10370702B2

## CPC

- C12Q 1/6811: Selection methods for production or design of target specific oligonucleotides or binding molecules
- C12Q 1/6813: Hybridisation assays
- 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

## Abstract

The present invention relates to methods of making linear nucleic acid probes using rolling circle amplification methods.

## Claims

### Claim 1

A plurality of nucleic acid probes in the form of a single stranded nucleic acid concatamer of repeating nucleic acid probe sequences wherein each nucleic acid probe sequence comprises
a polymerase recognition site at one end, a first nucleic acid sequence complementary to a target sequence, and a label addition site
at the other end,
and further comprising a hybridized oligonucleotide at the polymerase recognition site to form a hybrid that is recognizable by an endonuclease.

### Claim 2 (depends on 1)

The nucleic acid probes of claim 1 wherein the single stranded nucleic acid concatamer is DNA.

### Claim 3 (depends on 1)

The nucleic acid probes of claim 1 wherein the single stranded nucleic acid concatamer is RNA.

### Claim 4 (depends on 1)

The nucleic acid probes of claim 1 wherein the polymerase recognition site is an RNA polymerase recognition site.

### Claim 5 (depends on 1)

The nucleic acid probes of claim 1 wherein the polymerase recognition site is a DNA polymerase recognition site.

### Claim 6 (depends on 1)

The nucleic acid probes of claim 1 wherein the label addition site comprises a sequence complementary to a secondary probe including a label.

### Claim 7 (depends on 1)

The method of claim 1 wherein the hybridized oligonucleotide is DNA and the polymerase recognition site is DNA to produce a DNA/DNA hybrid recognizable by the endonuclease.

### Claim 8 (depends on 1)

The method of claim 1 wherein the hybridized oligonucleotide is DNA and the polymerase recognition site is RNA to produce a DNA/RNA hybrid recognizable by the endonuclease.

### Claim 9 (depends on 1)

The method of claim 1 wherein the hybridized oligonucleotide is RNA and the polymerase recognition site is DNA to produce a RNA/DNA hybrid recognizable by the endonuclease.

### Claim 10 (depends on 1)

The method of claim 1 wherein the hybridized oligonucleotide is RNA and the polymerase recognition site is RNA to produce a RNA/RNA hybrid recognizable by the endonuclease.