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EP0496483A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Process to prevent contamination by amplified nucleic acid sequence

Assignee: Integrated DNA Technologies Inc
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Filing: 1992-01-02
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Granted: —
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Expires: 2012-01-02
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Abstract

Abstract An improved method for eliminating the problem of contamination by amplified nucleic acid sequences in which novel primers or polynucleotide substrates are used that can be cleaved from the product, thereby preventing it from being further amplified.

Inventors

  • 👤 Joseph A. Walder
  • 👤 Roxanne Y. Walder

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-10-29 23:09
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/68; C12Q1/6844; C12Q1/6848

Claims Analysis

Total Claims: 19
Claim 1
Independent Claim 41 words
A process for decreasing contamination by an amplified product in nucleic acid amplification reactions, said process comprising: using a primer or polynucleotide substrate which contains a ribose residue in the amplification reaction; and cleaving the ribonucleotide linkage within the amplified product.
Claim 2
Dependent Claim 21 words
A process as claimed in claim 1, in which cleavage of the ribonucleotide linkage is performed prior to the amplification reaction.
Claim 3
Dependent Claim 20 words
A process as claimed in claim 1, in which cleavage of the ribonucleotide linkage is performed after the amplification reaction.
Claim 4
Dependent Claim 21 words
A process as claimed in claim 1, 2 or 3, in which said amplification reaction is the polymerase chain reaction (PCR).
Claim 5
Dependent Claim 21 words
A process as claimed in claim 1, 2 or 3, in which said amplification reaction is a transcription based amplification reaction.
Claim 6
Dependent Claim 20 words
A process as claimed in claim 1, 2 or 3, in which said amplification reaction is the ligase chain reaction.
Claim 7
Dependent Claim 20 words
A process as claimed in any preceding claim, in which said primer contains a single ribose residue at its 3′-terminus.
Claim 8
Dependent Claim 20 words
A process as claimed in any preceding claim, in which cleavage of the ribonucleotide linkage is accomplished using an enzyme.
Claim 9
Dependent Claim 19 words
A process as claimed in claim 8, wherein said enzyme is selected from RNaseIII, RNaseH and single stranded ribonucleases.
Claim 10
Dependent Claim 25 words
A process as claimed in any one of claims 1 to 9, in which cleavage of the ribonucleotide linkage is accomplished by treatment with base.
Claim 11
Dependent Claim 50 words
A process for isolating an amplified nucleic acid sequence, said process comprising: using a primer or polynucleotide substrate containing a ribose residue in the amplification reaction to produce said sequence; binding said sequence to a solid support, and releasing said sequence from said solid support by cleaving the ribonucleotide linkage.
Claim 12
Dependent Claim 21 words
A process as claimed in claim 11, in which said sequence is bound to a solid support through a biotin-avidin linkage.
Claim 13
Dependent Claim 19 words
A process as claimed in claim 11 or 12, in which the ribonucleotide linkage is cleaved using an enzyme.
Claim 14
Dependent Claim 16 words
A process as claimed in claim 13, in which said enzyme is a single stranded ribonuclease.
Claim 15
Dependent Claim 18 words
A process as claimed in claim 11 or 12, in which the ribonucleotide linkage is cleaved using base.
Claim 16
Dependent Claim 59 words
A kit for detection of a nucleic acid sequences which minimizes contamination by an amplified product comprising: a primer or polynucleotide substrate able to hybridize to a specific target nucleic acid sequence and containing a ribose residue that becomes incorporated into an amplification product; and a reagent able to cleave the ribonucleotide linkage of said primer or polynucleotide substrate.
Claim 17
Dependent Claim 17 words
A kit as claimed in claim 16, in which said primer contains a 3′ terminal ribonucleotide residue.
Claim 18
Dependent Claim 17 words
A kit as claimed in claim 16 or 17, in which said cleavage reagent is an enzyme.
Claim 19
Dependent Claim 18 words
A kit as claimed in claim 16 or 17, in which said cleavage reagent is a basic solution.

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