Emulsion PCR (ePCR) as a Tool to Improve the Power of DGGE Analysis for Microbial Population Studies.

Microorganisms

Department of Agricultural, Food, Environmental and Animal Science, University of Udine, via Sondrio 2/A, 33100 Udine, Italy.

Published: July 2020

To the authors' knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studies was tested. Our results indicated that ePCR results in uniform amplification of several DNA molecules, overcoming the major limitations of conventional PCR, such as preferential amplification and DNA concentration dependence. Moreover, ePCR-DGGE resulted in higher sensitivity when compared to conventional PCR-DGGE methods used for studying microbial populations in a complex matrix. In fact, compared to conventional PCR, the DGGE profiles of ePCR products permitted the detection of a higher number of the species that were present in the tested sample.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465085PMC
http://dx.doi.org/10.3390/microorganisms8081099DOI Listing

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