Cargando…

Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles

High-molecular-weight DNA (HMW DNA) extracted from soil is useful for examined the functions and diversity of soil organisms, the majority of which are difficult to culture. In the present study, the procedures used to extract HMW DNA from soil samples were improved. The grinding of soil samples wit...

Descripción completa

Detalles Bibliográficos
Autor principal: Sakai, Yoriko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8446751/
https://www.ncbi.nlm.nih.gov/pubmed/34234043
http://dx.doi.org/10.1264/jsme2.ME21017
_version_ 1784568953731809280
author Sakai, Yoriko
author_facet Sakai, Yoriko
author_sort Sakai, Yoriko
collection PubMed
description High-molecular-weight DNA (HMW DNA) extracted from soil is useful for examined the functions and diversity of soil organisms, the majority of which are difficult to culture. In the present study, the procedures used to extract HMW DNA from soil samples were improved. The grinding of soil samples with liquid nitrogen followed by a lysozyme treatment at 45°C for 1 h and an incubation with protease and SDS at 50°C for 5 h increased the size and yield of HMW DNA extracted from these samples. In the soil group Andosols, the addition of boiled sonicated salmon DNA was effective for HMW DNA extraction.
format Online
Article
Text
id pubmed-8446751
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles
record_format MEDLINE/PubMed
spelling pubmed-84467512021-09-29 Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles Sakai, Yoriko Microbes Environ Short Communication High-molecular-weight DNA (HMW DNA) extracted from soil is useful for examined the functions and diversity of soil organisms, the majority of which are difficult to culture. In the present study, the procedures used to extract HMW DNA from soil samples were improved. The grinding of soil samples with liquid nitrogen followed by a lysozyme treatment at 45°C for 1 h and an incubation with protease and SDS at 50°C for 5 h increased the size and yield of HMW DNA extracted from these samples. In the soil group Andosols, the addition of boiled sonicated salmon DNA was effective for HMW DNA extraction. Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles 2021 2021-07-06 /pmc/articles/PMC8446751/ /pubmed/34234043 http://dx.doi.org/10.1264/jsme2.ME21017 Text en 2021 by Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles. https://creativecommons.org/licenses/by/3.0/This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Short Communication
Sakai, Yoriko
Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
title Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
title_full Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
title_fullStr Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
title_full_unstemmed Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
title_short Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
title_sort improvements in extraction methods of high-molecular-weight dna from soils by modifying cell lysis conditions and reducing adsorption of dna onto soil particles
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8446751/
https://www.ncbi.nlm.nih.gov/pubmed/34234043
http://dx.doi.org/10.1264/jsme2.ME21017
work_keys_str_mv AT sakaiyoriko improvementsinextractionmethodsofhighmolecularweightdnafromsoilsbymodifyingcelllysisconditionsandreducingadsorptionofdnaontosoilparticles