Direct extraction of microbial community DNA from humified upland soils.
This paper describes a protocol effective at extracting high yields of high-purity microbial community DNA from humified soils. DNA was extracted from soil by lysozyme, SDS and freeze-thaw lysis, precipitated and then subjected to a double caesium chloride density gradient centrifugation stage before concentrating and washing. Evaluation using three soils yielded up to 30 micrograms DNA g-1 dry soil, with absorbance ratios at 260:230 nm and 260:280 nm of 1.6-2.0. The DNA extracted from the three soils was digested by four restriction enzymes and a 16S rDNA eubacterial product was amplified by PCR. These tests indicated that the DNA obtained by the protocol was sufficiently pure for molecular biological analysis.
Clegg
C D
CD
Cellular and Environmental Physiology Department, Scottish Crop Research Institute, Invergowrie, Dundee, UK.
Ritz
K
K
Griffiths
B S
BS
D004269:DNA, Bacterial / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012988:Soil Microbiology
D004269:DNA, Bacterial