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Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis

Genome analysis using pulsed-field gel electrophoresis (PFGE) has been used in applications ranging from typing bacterial strains to radiobiology to cancer research. While methods for running PFGE have been significantly improved since its invention, the method for preparing chromosomal DNA itself h...

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Detalles Bibliográficos
Autores principales: Hicks, Linda, van der Graaf, Charlotte M., Childress, Jacob, Cook, Emily, Schmidt, Karen, Rosenzweig, Frank, Kroll, Eugene
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Journal of Biological Methods 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6706164/
https://www.ncbi.nlm.nih.gov/pubmed/31453239
http://dx.doi.org/10.14440/jbm.2018.218
Descripción
Sumario:Genome analysis using pulsed-field gel electrophoresis (PFGE) has been used in applications ranging from typing bacterial strains to radiobiology to cancer research. While methods for running PFGE have been significantly improved since its invention, the method for preparing chromosomal DNA itself has remained essentially unchanged. This limits the applicability of PFGE, especially when analyses require many samples. We have streamlined sample preparation for routine applications of PFGE through the use of deep-well 48-well plates. Besides saving time, our protocol has the added advantage of reducing the volume of expensive reagents. Our improved protocol enables us to reduce throughput time and simplify the procedure, facilitating wider application of PFGE-based analyses in the laboratory.