# US 2010/0167353 A1 · Rnase h-based assays utilizing modified rna monomers

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- Number: US 2010/0167353 A1
- Document ID: US20100167353A1
- Status: Published application
- Kind: A1
- Application number: 12/507,142
- Filed: 2009-07-22
- Published: 2010-07-01
- Earliest claimed: 2008-04-30
- Assignees: Integrated Dna Technologies, Inc.
- Inventors: Joseph Alan Walder; Mark Aaron Behlke; Scott Rose; Joseph Dobosy
- Web page: https://patentgrep.com/patent/US20100167353A1

## CPC

- C12Q 1/6848: characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
- C12Q 1/6844: Nucleic acid amplification reactions
- C12Q 1/6853: using modified primers or templates
- C12Q 1/6858: Allele-specific amplification
- 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

## Abstract

The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3′ end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.

## Claims

### Claim 1

A method of amplifying a target DNA sequence, said method comprising the steps of:
a) providing a reaction mixture comprising (i) an oligonucleotide primer having a cleavage domain positioned 5′ of a blocking group, said blocking group linked at or near the end of the 3′-end of the oligonucleotide primer wherein said blocking group prevents primer extension, (ii) a sample nucleic acid that may or may not have the target sequence, (iii) a cleaving enzyme and (iv) a polymerase wherein said cleaving enzyme is a hot start cleaving enzyme which is thermostable and has reduced activity at lower temperatures;
b) hybridizing the primer to the target DNA sequence to form a double-stranded substrate;
c) cleaving the hybridized primer with said cleaving enzyme at a point within or adjacent to the cleavage domain to remove the blocking group from the primer; and extending the primer with the polymerase.

### Claim at position 2

2-140. (canceled)