Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Zaveri, Purvi, Patel, Rushika, Patel, Meghavi, Sarodia, Devki, Munshi, Nasreen S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
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
_version_ 1782505449262678016
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
work_keys_str_mv AT zaveripurvi modificationofextractionmethodforcommunitydnaisolationfromsaltaffectedcompactwastelandsoilsamples
AT patelrushika modificationofextractionmethodforcommunitydnaisolationfromsaltaffectedcompactwastelandsoilsamples
AT patelmeghavi modificationofextractionmethodforcommunitydnaisolationfromsaltaffectedcompactwastelandsoilsamples
AT sarodiadevki modificationofextractionmethodforcommunitydnaisolationfromsaltaffectedcompactwastelandsoilsamples
AT munshinasreens modificationofextractionmethodforcommunitydnaisolationfromsaltaffectedcompactwastelandsoilsamples