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
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 18:04
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.
References Cited (Backward Citations)
No citation records indexed.
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