Improvements in Extraction Methods of High-molecular-weight DNA from Soils by Modifying Cell Lysis Conditions and Reducing Adsorption of DNA onto Soil Particles
Yoriko Sakai
Abstract
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.
Topics & Concepts
DNA extractionLysisDNASoil waterExtraction (chemistry)ChemistrySonicationLysozymeIncubationAdsorptionLysis bufferChromatographySoil testBiologyBiochemistryPolymerase chain reactionGeneEcologyOrganic chemistryEnvironmental DNA in Biodiversity StudiesMicrobial Community Ecology and PhysiologyGenomics and Phylogenetic Studies