Direct extraction of DNA from soils for studies in microbial ecology

dc.contributor.authorSchneegurt, Mark A.en_US
dc.contributor.authorDore, Sophia Y.en_US
dc.contributor.authorKulpa, Charles F.en_US
dc.descriptionClick on the link below to access the article (may not be free).en_US
dc.description.abstractMolecular analyses for the study of soil microbial communities often depend on the extraction of DNA directly from soils. These extractions are by no means trivial, being complicated by humic substances that are inhibitory to PCR and restriction enzymes or being too highly colored for blot hybridization protocols. Many different published protocols exist, but none have been found to be suitable enough to be generally accepted as a standard. Most direct extraction protocols start with relatively harsh cell breakage steps such as bead-beating and freeze-thaw cycles, followed by the addition of detergents and high salt buffers and/or enzymic digestion with lysozyme and proteases. After typical organic extraction and alcohol precipitation, further purification is usually needed to remove inhibitory substances from the extract. The purification steps include size-exclusion chromatography, ion-exchange chromatography, silica gel spin columns, and cesium chloride gradients, among others. A direct DNA extraction protocol is described that has been shown to be effective in a wide variety of soil types. This protocol is experimentally compared to several published protocols.en_US
dc.description.versionpeer revieweden_US
dc.identifier.citationCurrent issues in molecular biology. 2003 Jan; 5(1): 1-8.en_US
dc.publisherHorizon Scientific Pressen_US
dc.relation.ispartofseriesCurrent issues in molecular biologyen_US
dc.rights.holderCopyright © 2003 Caister Academic Pressen_US
dc.subjectResearch Support, Non-U.S. Gov'ten_US
dc.subject.meshDNA/isolation and purificationen_US
dc.subject.meshDNA, Ribosomal/analysisen_US
dc.subject.meshGenetic Techniquesen_US
dc.subject.meshSoil Microbiologyen_US
dc.subject.meshDNA, Ribosomal/isolation & purificationen_US
dc.titleDirect extraction of DNA from soils for studies in microbial ecologyen_US