Evaluation of quantitative polymerase chain reaction to assess nosZ gene prevalence in mixed microbial communities.
The usefulness of quantitative polymerase chain reaction (QPCR) to measure nosZ gene prevalence in a multi-template reaction was assessed by comparing 19 nosZ template DNA samples and 91 model communities. Efficiencies of the QPCR varied but were not significantly different among nosZ genotypes and were not linked to genetic distance from Ralstonia eutropha. nosZ genotype QPCR efficiencies obtained from isolated denitrifiers were higher (84.8%) than those obtained from excised denaturing gradient gel electrophoresis bands or clones of PCR products from total community DNA (ca. 60%). Analysis of the model communities indicated that QPCR accurately predicts gene prevalence in communities composed of up to six templates.
Siciliano
Steven D
SD
Department of Soil Science, University of Saskatchewan, Saskatoon, SK S7N 5A8, Canada. steven.siciliano@usask.ca
Ma
Wai
W
Powell
Shane
S
D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004463:Ecology; D005069:Evaluation Studies as Topic; D005798:Genes, Bacterial; D010088:Oxidoreductases / Q000032:analysis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012988:Soil Microbiology
D004269:DNA, Bacterial; D010088:Oxidoreductases; C034747:nitric-oxide reductase