Rapid, direct extraction of DNA from soils for PCR analysis using polyvinylpolypyrrolidone spin columns.
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.
Berthelet
M
M
National Research Council of Canada, Biotechnology Research Institute, Montréal, Québec, Canada.
Whyte
L G
LG
Greer
C W
CW
D015084:2,4-Dichlorophenoxyacetic Acid / Q000378:metabolism; D001419:Bacteria / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001673:Biodegradation, Environmental; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D049308:Dioxygenases; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011205:Povidone / Q000031:analogs & derivatives; D012988:Soil Microbiology; D012989:Soil Pollutants / Q000378:metabolism
D017931:DNA Primers; D004269:DNA, Bacterial; D012989:Soil Pollutants; C077842:polyvinylpolypyrrolidone; D015084:2,4-Dichlorophenoxyacetic Acid; D010105:Oxygenases; D049308:Dioxygenases; C053101:catechol oxygenase II; D011205:Povidone