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Pitfalls encountered while investigating genetic elements by PCR

The unprecedented wealth of databases that have become available in the era of next-generation sequencing has considerably increased our knowledge of bacterial genetic elements (GEs). At the same time, the advent of single-cell based approaches has brought realization that unsuspected heterogeneity...

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Detalles Bibliográficos
Autores principales: Palmieri, Claudio, Brenciani, Andrea, Mingoia, Marina, Giovanetti, Eleonora, Facinelli, Bruna, Varaldo, Pietro E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Landes Bioscience 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3742598/
https://www.ncbi.nlm.nih.gov/pubmed/23956950
http://dx.doi.org/10.4161/mge.25255
Descripción
Sumario:The unprecedented wealth of databases that have become available in the era of next-generation sequencing has considerably increased our knowledge of bacterial genetic elements (GEs). At the same time, the advent of single-cell based approaches has brought realization that unsuspected heterogeneity may occur in the bacterial population from a single colony. The increasing use of PCR-based techniques to study new GEs requires careful consideration of the possible different PCR targets associated with different subpopulations if incorrect or incomplete interpretations are to be avoided. In this commentary, confining ourselves to our direct experience, we illustrate some examples of PCR pitfalls that may be encountered while investigating GEs.