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Cellular reagents for diagnostics and synthetic biology

We have found that the overproduction of enzymes in bacteria followed by their lyophilization leads to 'cellular reagents' that can be directly used to carry out numerous molecular biology reactions. We demonstrate the use of cellular reagents in a variety of molecular diagnostics, such as...

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Detalles Bibliográficos
Autores principales: Bhadra, Sanchita, Pothukuchy, Arti, Shroff, Raghav, Cole, Austin W., Byrom, Michelle, Ellefson, Jared W., Gollihar, Jimmy D., Ellington, Andrew D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6093680/
https://www.ncbi.nlm.nih.gov/pubmed/30110361
http://dx.doi.org/10.1371/journal.pone.0201681
Descripción
Sumario:We have found that the overproduction of enzymes in bacteria followed by their lyophilization leads to 'cellular reagents' that can be directly used to carry out numerous molecular biology reactions. We demonstrate the use of cellular reagents in a variety of molecular diagnostics, such as TaqMan qPCR with no diminution in sensitivity, and in synthetic biology cornerstones such as the Gibson assembly of DNA fragments, where new plasmids can be constructed solely based on adding cellular reagents. Cellular reagents have significantly reduced complexity and cost of production, storage and implementation, features that should facilitate accessibility and use in resource-poor conditions.