Extraction and purification of DNA in rhizosphere soil samples for PCR-DGGE analysis of bacterial consortia.
Application of DNA fingerprinting methods enables the detection of diverse members of soil bacterial consortia, even including those bacteria not yet cultivated. However, extraction and purification of DNA from soil samples without bias is difficult. We compared five different DNA isolation methods and three purification methods for rhizosphere soil samples. Purified DNA extracts were amplified in PCR using universal bacterial primers and the PCR products were analysed with denaturing gradient gel electrophoresis (DGGE) for the visualisation of DNA bands representing dominant bacterial species. Both the isolation and purification methods affected the apparent bacterial community structure of the samples.
Maarit Niemi
R
R
Finnish Environment Institute, PO Box 140 FIN-00251, Helsinki, Finland. Maarit.niemi@vyh.fi
Heiskanen
I
I
Wallenius
K
K
Lindström
K
K
D001419:Bacteria / Q000235:genetics*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012988:Soil Microbiology
D004269:DNA, Bacterial