Iacumin Lucilla, Cecchini Francesca, Vendrame Marco, Comi Giuseppe
Department of Agricultural, Food, Environmental and Animal Science, University of Udine, via Sondrio 2/A, 33100 Udine, Italy.
Microorganisms. 2020 Jul 23;8(8):1099. doi: 10.3390/microorganisms8081099.
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.
据作者所知，这是关于乳液聚合酶链反应（e-PCR）与变性梯度凝胶电泳（DGGE）分析联用的首次报道。在本研究中，测试了ePCR在提高DGGE技术用于微生物群体研究效能方面的有效性。我们的结果表明，ePCR能使多个DNA分子实现均匀扩增，克服了传统PCR的主要局限性，如优先扩增和对DNA浓度的依赖性。此外，与用于研究复杂基质中微生物群体的传统PCR-DGGE方法相比，ePCR-DGGE具有更高的灵敏度。事实上，与传统PCR相比，ePCR产物的DGGE图谱能够检测到测试样品中存在的更多物种。
