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Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples
To overcome the issue of interferences by salt and compactness in release of bacterial cell required for lysis, method described by Yeates et al. (1998), was optimized for isolation of genomic material (Deoxyribo Nucleic Acid, DNA) from soil microbial community by addition of Al(NH(4))SO(4). Very lo...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5295508/ https://www.ncbi.nlm.nih.gov/pubmed/28203533 http://dx.doi.org/10.1016/j.mex.2017.01.002 |
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author | Zaveri, Purvi Patel, Rushika Patel, Meghavi Sarodia, Devki Munshi, Nasreen S. |
author_facet | Zaveri, Purvi Patel, Rushika Patel, Meghavi Sarodia, Devki Munshi, Nasreen S. |
author_sort | Zaveri, Purvi |
collection | PubMed |
description | To overcome the issue of interferences by salt and compactness in release of bacterial cell required for lysis, method described by Yeates et al. (1998), was optimized for isolation of genomic material (Deoxyribo Nucleic Acid, DNA) from soil microbial community by addition of Al(NH(4))SO(4). Very low total viable count was observed in the samples tested and hence use of higher amount of soil is required primarily for DNA isolation from wasteland soils. The method proves itself efficient where commercially available bead beating and enzymatic lysis methods could not give isolation of any amount of community genomic DNA due to compact nature and salt concentrations present in soil. • The protocol was found efficient for soil samples with high clay content for microbial community DNA extraction. • Variation in lysis incubation and amount of soil may help with soil samples containing low microbial population. • Addition of Al(NH(4))SO(4) is crucial step in humic acid removal from extracted DNA samples for soil samples containing high salinity and clay particles. |
format | Online Article Text |
id | pubmed-5295508 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-52955082017-02-15 Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples Zaveri, Purvi Patel, Rushika Patel, Meghavi Sarodia, Devki Munshi, Nasreen S. MethodsX Environmental Science To overcome the issue of interferences by salt and compactness in release of bacterial cell required for lysis, method described by Yeates et al. (1998), was optimized for isolation of genomic material (Deoxyribo Nucleic Acid, DNA) from soil microbial community by addition of Al(NH(4))SO(4). Very low total viable count was observed in the samples tested and hence use of higher amount of soil is required primarily for DNA isolation from wasteland soils. The method proves itself efficient where commercially available bead beating and enzymatic lysis methods could not give isolation of any amount of community genomic DNA due to compact nature and salt concentrations present in soil. • The protocol was found efficient for soil samples with high clay content for microbial community DNA extraction. • Variation in lysis incubation and amount of soil may help with soil samples containing low microbial population. • Addition of Al(NH(4))SO(4) is crucial step in humic acid removal from extracted DNA samples for soil samples containing high salinity and clay particles. Elsevier 2017-01-17 /pmc/articles/PMC5295508/ /pubmed/28203533 http://dx.doi.org/10.1016/j.mex.2017.01.002 Text en © 2017 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Environmental Science Zaveri, Purvi Patel, Rushika Patel, Meghavi Sarodia, Devki Munshi, Nasreen S. Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples |
title | Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples |
title_full | Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples |
title_fullStr | Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples |
title_full_unstemmed | Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples |
title_short | Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples |
title_sort | modification of extraction method for community dna isolation from salt affected compact wasteland soil samples |
topic | Environmental Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5295508/ https://www.ncbi.nlm.nih.gov/pubmed/28203533 http://dx.doi.org/10.1016/j.mex.2017.01.002 |
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