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SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol

Direct isolation of soil DNA comes as an emerging technology to understand the microbial diversity of a particular environment circumventing the dependency on culturable methods. Soil DNA isolation is tough due to the presence of various organic components present in soil which interfere in extracti...

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Detalles Bibliográficos
Autores principales: Paul, Tanima, Basu, Semanti, Sarkar, Keka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235891/
https://www.ncbi.nlm.nih.gov/pubmed/28324313
http://dx.doi.org/10.1007/s13205-014-0232-y
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author Paul, Tanima
Basu, Semanti
Sarkar, Keka
author_facet Paul, Tanima
Basu, Semanti
Sarkar, Keka
author_sort Paul, Tanima
collection PubMed
description Direct isolation of soil DNA comes as an emerging technology to understand the microbial diversity of a particular environment circumventing the dependency on culturable methods. Soil DNA isolation is tough due to the presence of various organic components present in soil which interfere in extraction procedure. Here, we report a novel direct soil DNA extraction protocol utilizing bare superparamagnetic iron oxide nanoparticles and its comparison with conventional and commercial kit-based soil DNA extraction methods. The quality, quantity and feasibility of the recovered DNA from all the three methods towards various molecular techniques were checked. Our magnetic nanoparticle-based soil DNA extraction successfully yields pure DNA without any RNA or protein contamination as revealed by the nanodrop spectrophotometer and agarose gel electrophoretic study. Different methods of soil DNA extraction were evaluated on the basis of PCR, denaturing gradient gel electrophoresis and real-time PCR. Soil DNA extracted using conventional method fails to carry out critical molecular biology techniques where as magnetic nanoparticle-based soil DNA extraction gave good results which is comparable to commercial kit. This comparative study suggests that protocol described in this report is novel, less time consuming, cost effective with fewer handling steps and yields high quantity, good quality DNA from soil.
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spelling pubmed-42358912014-11-20 SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol Paul, Tanima Basu, Semanti Sarkar, Keka 3 Biotech Protocols and Methods Direct isolation of soil DNA comes as an emerging technology to understand the microbial diversity of a particular environment circumventing the dependency on culturable methods. Soil DNA isolation is tough due to the presence of various organic components present in soil which interfere in extraction procedure. Here, we report a novel direct soil DNA extraction protocol utilizing bare superparamagnetic iron oxide nanoparticles and its comparison with conventional and commercial kit-based soil DNA extraction methods. The quality, quantity and feasibility of the recovered DNA from all the three methods towards various molecular techniques were checked. Our magnetic nanoparticle-based soil DNA extraction successfully yields pure DNA without any RNA or protein contamination as revealed by the nanodrop spectrophotometer and agarose gel electrophoretic study. Different methods of soil DNA extraction were evaluated on the basis of PCR, denaturing gradient gel electrophoresis and real-time PCR. Soil DNA extracted using conventional method fails to carry out critical molecular biology techniques where as magnetic nanoparticle-based soil DNA extraction gave good results which is comparable to commercial kit. This comparative study suggests that protocol described in this report is novel, less time consuming, cost effective with fewer handling steps and yields high quantity, good quality DNA from soil. Springer Berlin Heidelberg 2014-06-24 2014-12 /pmc/articles/PMC4235891/ /pubmed/28324313 http://dx.doi.org/10.1007/s13205-014-0232-y Text en © The Author(s) 2014 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Protocols and Methods
Paul, Tanima
Basu, Semanti
Sarkar, Keka
SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol
title SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol
title_full SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol
title_fullStr SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol
title_full_unstemmed SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol
title_short SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol
title_sort spion-mediated soil dna extraction and comparative analysis with conventional and commercial kit-based protocol
topic Protocols and Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235891/
https://www.ncbi.nlm.nih.gov/pubmed/28324313
http://dx.doi.org/10.1007/s13205-014-0232-y
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