Applied and Environmental Microbiology · 1991 · 622 citations · 19 references
AgricultureMicrobiologySoil SciencesSoil BiologySoil MicrobiologySoil ScienceNucleic Acid ExtractionNucleic Acid PurificationSediment MicrobiologyDna ExtractionEnvironmental Dna ExtractionMicrobial Nucleic Acid ExtractionRecovery EfficiencySedimentary DnaShearingSodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis
A rapid method for the direct extraction of DNA from soil and sediments was developed. The indigenous microorganisms in the soil and sediments were lysed by using lysozyme and a freeze-thaw procedure. The lysate was extracted with sodium dodecyl sulfate and phenol-chloroform. In addition to a high recovery efficiency (greater than 90%), the yields of DNA were high (38 and 12 micrograms/g [wet weight] from sediments and soil, respectively). This method generated minimal shearing of the extracted DNA. The crude DNA could be further purified with an Elutip-d column if necessary. An additional advantage of this method is that only 1 g of sample is required, which allows for the analysis of small samples and the processing of many samples in a relatively short (7 h) period.
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Recovery of DNA from soils and sediments
Robert J. Steffan, Jostein Goksøyr, A K Bej et al. · Applied and Environmental Microbiology · 1988 · 485 citations · Full text