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Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies

To the authors’ knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studi...

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Detalles Bibliográficos
Autores principales: Iacumin, Lucilla, Cecchini, Francesca, Vendrame, Marco, Comi, Giuseppe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465085/
https://www.ncbi.nlm.nih.gov/pubmed/32717823
http://dx.doi.org/10.3390/microorganisms8081099
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author Iacumin, Lucilla
Cecchini, Francesca
Vendrame, Marco
Comi, Giuseppe
author_facet Iacumin, Lucilla
Cecchini, Francesca
Vendrame, Marco
Comi, Giuseppe
author_sort Iacumin, Lucilla
collection PubMed
description To the authors’ knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studies was tested. Our results indicated that ePCR results in uniform amplification of several DNA molecules, overcoming the major limitations of conventional PCR, such as preferential amplification and DNA concentration dependence. Moreover, ePCR-DGGE resulted in higher sensitivity when compared to conventional PCR-DGGE methods used for studying microbial populations in a complex matrix. In fact, compared to conventional PCR, the DGGE profiles of ePCR products permitted the detection of a higher number of the species that were present in the tested sample.
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spelling pubmed-74650852020-09-04 Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies Iacumin, Lucilla Cecchini, Francesca Vendrame, Marco Comi, Giuseppe Microorganisms Article To the authors’ knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studies was tested. Our results indicated that ePCR results in uniform amplification of several DNA molecules, overcoming the major limitations of conventional PCR, such as preferential amplification and DNA concentration dependence. Moreover, ePCR-DGGE resulted in higher sensitivity when compared to conventional PCR-DGGE methods used for studying microbial populations in a complex matrix. In fact, compared to conventional PCR, the DGGE profiles of ePCR products permitted the detection of a higher number of the species that were present in the tested sample. MDPI 2020-07-23 /pmc/articles/PMC7465085/ /pubmed/32717823 http://dx.doi.org/10.3390/microorganisms8081099 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Iacumin, Lucilla
Cecchini, Francesca
Vendrame, Marco
Comi, Giuseppe
Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
title Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
title_full Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
title_fullStr Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
title_full_unstemmed Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
title_short Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies
title_sort emulsion pcr (epcr) as a tool to improve the power of dgge analysis for microbial population studies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465085/
https://www.ncbi.nlm.nih.gov/pubmed/32717823
http://dx.doi.org/10.3390/microorganisms8081099
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