Open AccessRapid, direct extraction of DNA from soils for PCR analysis using polyvinylpolypyrrolidone spin columns
Author(s) -
Berthelet Marc,
Whyte Lyle G.,
Greer Charles W.
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08128.x
Subject(s) - polymerase chain reaction , dna extraction , soil water , dna , extraction (chemistry) , chromatography , chemistry , biology , soil microbiology , detection limit , gene , microbiology and biotechnology , biochemistry , ecology
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.