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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...

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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
Descripción
Sumario: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.