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The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices

Molecular approaches are powerful tools that are used for medical or environmental diagnoses. However, the main limitations of such a tools are that they extract low levels of DNA and they do not remove the inhibitors of polymerase chain reaction (PCR). Although the use of polycation to complex and...

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Autores principales: François, Celia, Martinez, Celia, Faye, Clement, Pansu, Nathalie, Dunyach-Remy, Catherine, Garrelly, Laurent, Roig, Benoit, Cadiere, Axelle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760326/
https://www.ncbi.nlm.nih.gov/pubmed/33266082
http://dx.doi.org/10.3390/microorganisms8121901
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author François, Celia
Martinez, Celia
Faye, Clement
Pansu, Nathalie
Dunyach-Remy, Catherine
Garrelly, Laurent
Roig, Benoit
Cadiere, Axelle
author_facet François, Celia
Martinez, Celia
Faye, Clement
Pansu, Nathalie
Dunyach-Remy, Catherine
Garrelly, Laurent
Roig, Benoit
Cadiere, Axelle
author_sort François, Celia
collection PubMed
description Molecular approaches are powerful tools that are used for medical or environmental diagnoses. However, the main limitations of such a tools are that they extract low levels of DNA and they do not remove the inhibitors of polymerase chain reaction (PCR). Although the use of polycation to complex and purify DNA has been described in the literature, elution often requires a high ionic strength or pH levels not compatible with molecular analyses. In this paper, we described a new process that is based on the complexation of DNA with linear polylysine, followed by capturing the complex by a cation exchange resin. The originality of the process consisted of using mechanic force to elute DNA from the complex. The extraction method showed several advantages when compared to existing methods, such as being compatible with pH levels that range from 5 to 11, as well as high levels of DNA recovery and elimination of PCR inhibitors from complex samples. This method was successfully applied to different types of samples, such as environmental samples, beverage samples, and medical samples. Furthermore, it was proven to be a good solution for removing PCR inhibitors and assuring good DNA recovery yield.
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spelling pubmed-77603262020-12-26 The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices François, Celia Martinez, Celia Faye, Clement Pansu, Nathalie Dunyach-Remy, Catherine Garrelly, Laurent Roig, Benoit Cadiere, Axelle Microorganisms Communication Molecular approaches are powerful tools that are used for medical or environmental diagnoses. However, the main limitations of such a tools are that they extract low levels of DNA and they do not remove the inhibitors of polymerase chain reaction (PCR). Although the use of polycation to complex and purify DNA has been described in the literature, elution often requires a high ionic strength or pH levels not compatible with molecular analyses. In this paper, we described a new process that is based on the complexation of DNA with linear polylysine, followed by capturing the complex by a cation exchange resin. The originality of the process consisted of using mechanic force to elute DNA from the complex. The extraction method showed several advantages when compared to existing methods, such as being compatible with pH levels that range from 5 to 11, as well as high levels of DNA recovery and elimination of PCR inhibitors from complex samples. This method was successfully applied to different types of samples, such as environmental samples, beverage samples, and medical samples. Furthermore, it was proven to be a good solution for removing PCR inhibitors and assuring good DNA recovery yield. MDPI 2020-11-30 /pmc/articles/PMC7760326/ /pubmed/33266082 http://dx.doi.org/10.3390/microorganisms8121901 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 Communication
François, Celia
Martinez, Celia
Faye, Clement
Pansu, Nathalie
Dunyach-Remy, Catherine
Garrelly, Laurent
Roig, Benoit
Cadiere, Axelle
The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices
title The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices
title_full The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices
title_fullStr The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices
title_full_unstemmed The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices
title_short The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices
title_sort utilization of linear polylysine coupled with mechanic forces to extract microbial dna from different matrices
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760326/
https://www.ncbi.nlm.nih.gov/pubmed/33266082
http://dx.doi.org/10.3390/microorganisms8121901
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