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Amplification of bacterial DNA bound on clay minerals by the random amplified polymorphic DNA (RAPD) technique
Authors:C Vettori  D Paffetti  G Pietramellara  G Stotzky  E Gallori
Institution:Department of Animal Biology and Genetics, University of Florence, via Romana 17, 50125 Florence, Italy;Department of Soil Science, University of Florence, Florence, Italy;Laboratory of Microbial Ecology, Department of Biology, New York University, New York, USA
Abstract:Abstract: Chromosomal DNA from Bacillus subtilis , bound on the clay minerals, montmorillonite (Wyoming (W) and Apache County (Ap)) and kaolinite (K), was subjected to the random amplified polymorphic DNA (RAPD) technique. DNA bound on the clays was not amplified with 0.625, 1.875, 6.25, and 12.5 U of Taq DNA polymerase, but amplification occurred when the clay-DNA complexes were diluted 10- and 20-fold or when 21 U of Taq DNA polymerase was added. DNA desorbed from the Ap-DNA and K-DNA equilibrium complexes was amplified with 0.625 U of Taq DNA polymerase, whereas amplification of DNA desorbed from the W-DNA complex occurred only after a 10-fold dilution or when 1.875 U of Taq DNA polymerase was used. These observations indicate that clay minerals differentially affect the amplification process, probably by inhibiting the activity of Taq DNA polymerase.
Keywords:RAPD  Environmental DNA  Clay-DNA complex
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