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Rapid, direct extraction of DNA from soils for PCR analysis using polyvinylpolypyrrolidone spin columns
M Berthelet1, L G Whyte, C W Greer
1National Research Council of Canada, Biotechnology Research Institute, Montréal, Québec, Canada.
FEMS Microbiology Letters
|April 15, 1996
Abstract:
Polyvinylpolypyrrolidone spin columns were used to rapidly purify crude soil DNA extracts from humic materials for polymerase chain reaction (PCR) analysis. The PCR detection limit for the tfdC gene, encoding chlorocatechol dioxygenase from the 2,4-dichlorophenoxyacetic acid degradation pathway, was 10(1)-10(2) cells/g soil in inoculated soils. The procedure could be applied to the amplification of biodegradative genes from indigenous microbial populations from a wide variety of soil types, and the entire analysis could be performed within 8 h.