Methods for DNA extraction from various soils: a comparison

Citation
Cc. Tien et al., Methods for DNA extraction from various soils: a comparison, J APPL MICR, 86(6), 1999, pp. 937-943
Citations number
23
Categorie Soggetti
Biology,Microbiology
Journal title
JOURNAL OF APPLIED MICROBIOLOGY
ISSN journal
13645072 → ACNP
Volume
86
Issue
6
Year of publication
1999
Pages
937 - 943
Database
ISI
SICI code
1364-5072(199906)86:6<937:MFDEFV>2.0.ZU;2-X
Abstract
Seven methods for bacterial DNA extraction and purification from soil sampl es were compared. Holben's direct lysis method recovered significantly grea ter amounts of DNA than the other methods tested, while CsCl-ethidium bromi de density gradient ultracentrifugation was better than gel filtration at r emoving humic acid from crude DNA isolated from soil. When both these metho ds were combined, 5.94 mu g of DNA (A(260/280) ratio around 1.754) was yiel ded g(-1) oven-dried sandstone shale alluvial soil; similarly satisfactory yields were obtained from Taiwan clay, and sandstone shale and slate alluvi al soil managed under different farming practices. DNA obtained by these me thods was readily digested by EcoR I and Hind III. When soil samples were s tored for 3 weeks at 4 degrees C, the fraction of high-molecular-weight DNA was reduced significantly. Thus, DNA extraction should be carried out as s oon as possible after a soil sample has been collected from the field. When hyphae of Pythium aphanidermatum and Fusarium solani were subjected to the above lysis method, DNA could not be detected in the extract.