Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube

A quantitative multiple competitive PCR (QMC-PCR) for determination of DNA copy numbers is described. Four competitive DNA templates for the env region of HIV-1 were constructed with sizes longer (187 and 163 bp) or shorter (122 and 105 bp) than the 142 bp of the wild-type PCR product. Varying amoun...

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Main Authors: Klaus Zimmermann, Daniela Schögl, Barbara Plaimauer, Josef W. Mannhalter
Format: Article
Language:English
Published: Taylor & Francis Group 1996-09-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/96213st06
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author Klaus Zimmermann
Daniela Schögl
Barbara Plaimauer
Josef W. Mannhalter
author_facet Klaus Zimmermann
Daniela Schögl
Barbara Plaimauer
Josef W. Mannhalter
author_sort Klaus Zimmermann
collection DOAJ
description A quantitative multiple competitive PCR (QMC-PCR) for determination of DNA copy numbers is described. Four competitive DNA templates for the env region of HIV-1 were constructed with sizes longer (187 and 163 bp) or shorter (122 and 105 bp) than the 142 bp of the wild-type PCR product. Varying amounts of each of these competitors are introduced together with the sample into a single reaction tube. Since competitors and wild-type fragments share the same primer recognition sequence (SK68/SK69), amplification occurs according to the rate of the introduced copy numbers. The PCR products are run on an agarose gel, and the copy number of the sample is determined by analyzing the bands with a video densitometer and calculating the equivalence point in a linear regression plot.
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spelling doaj-art-7e0c4210cecf4ca28b38d5ff180e90ad2025-08-20T02:26:06ZengTaylor & Francis GroupBioTechniques0736-62051940-98181996-09-0121348048410.2144/96213st06Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction TubeKlaus Zimmermann0Daniela Schögl1Barbara Plaimauer2Josef W. Mannhalter31Immuno AG, Vienna, Austria1Immuno AG, Vienna, Austria1Immuno AG, Vienna, Austria1Immuno AG, Vienna, AustriaA quantitative multiple competitive PCR (QMC-PCR) for determination of DNA copy numbers is described. Four competitive DNA templates for the env region of HIV-1 were constructed with sizes longer (187 and 163 bp) or shorter (122 and 105 bp) than the 142 bp of the wild-type PCR product. Varying amounts of each of these competitors are introduced together with the sample into a single reaction tube. Since competitors and wild-type fragments share the same primer recognition sequence (SK68/SK69), amplification occurs according to the rate of the introduced copy numbers. The PCR products are run on an agarose gel, and the copy number of the sample is determined by analyzing the bands with a video densitometer and calculating the equivalence point in a linear regression plot.https://www.future-science.com/doi/10.2144/96213st06
spellingShingle Klaus Zimmermann
Daniela Schögl
Barbara Plaimauer
Josef W. Mannhalter
Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube
BioTechniques
title Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube
title_full Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube
title_fullStr Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube
title_full_unstemmed Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube
title_short Quantitative Multiple Competitive PCR of HIV-1 DNA in a Single Reaction Tube
title_sort quantitative multiple competitive pcr of hiv 1 dna in a single reaction tube
url https://www.future-science.com/doi/10.2144/96213st06
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AT danielaschogl quantitativemultiplecompetitivepcrofhiv1dnainasinglereactiontube
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AT josefwmannhalter quantitativemultiplecompetitivepcrofhiv1dnainasinglereactiontube