Error estimation in environmental DNA targets quantification due to PCR efficiencies differences between real samples and standards
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Nowadays, as in the past, our knowledge of microbiology is limited because of the capacity of available technology tools. In recent decades, molecular techniques are playing a central role in the understanding of the microbial world. In this sense, a complete scientific evaluation of biological samples is not possible without using molecular biology. The most influential technique has probably been the polymerase chain reaction (PCR), which allows us to work beyond classical microbiology. With PCR, in some cases, the need for culture may be avoided. Nevertheless, until the development of real-time PCR, our evaluation of the microbial complexity has been merely qualitative.
Real-time PCR is an evolution of the conventional endpoint PCR wherein the amplification of a target gene and its fluorescence detection occurs simultaneously during each cycle. Different strategies based on the use of nonspecific DNA intercalating dyes or specific fluorescent probes exist for linking...
KeywordsPolymerase Chain Reaction Environmental Sample Polymerase Chain Reaction Reaction Polymerase Chain Reaction Efficiency Sample Polymerase Chain Reaction
We thank the Ministry of Education and Science of Spain and the Comissionat per a Universitats i Recerca del Departament d’Innovacio´ and Universitats i Empresa de la Generalitat de Catalunya i del Fons Social Europeu, for supporting this study. Financial support was provided by Alfa Network TECSPAR (RED ALFA II-0543- FI-FAFCD; Sustainable technologies for potabilization and wastewater treatment), and by grant CTM2008-06676-C05-02/TECNO from the Ministry of Science and Innovation of Spain to Jordi Morató. The authors also want to thank to the peer review process of Folia Microbiologica for improving the quality of the present work.
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