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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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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
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author Hicks, Linda
van der Graaf, Charlotte M.
Childress, Jacob
Cook, Emily
Schmidt, Karen
Rosenzweig, Frank
Kroll, Eugene
author_facet Hicks, Linda
van der Graaf, Charlotte M.
Childress, Jacob
Cook, Emily
Schmidt, Karen
Rosenzweig, Frank
Kroll, Eugene
author_sort Hicks, Linda
collection PubMed
description 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.
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spelling pubmed-67061642019-08-26 Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis Hicks, Linda van der Graaf, Charlotte M. Childress, Jacob Cook, Emily Schmidt, Karen Rosenzweig, Frank Kroll, Eugene J Biol Methods Protocol 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. Journal of Biological Methods 2018-03-09 /pmc/articles/PMC6706164/ /pubmed/31453239 http://dx.doi.org/10.14440/jbm.2018.218 Text en © 2013-2018 The Journal of Biological Methods, All rights reserved. https://creativecommons.org/licenses/by/3.0/ This work is licensed under a Creative Commons Attribution 3.0 License.
spellingShingle Protocol
Hicks, Linda
van der Graaf, Charlotte M.
Childress, Jacob
Cook, Emily
Schmidt, Karen
Rosenzweig, Frank
Kroll, Eugene
Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
title Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
title_full Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
title_fullStr Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
title_full_unstemmed Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
title_short Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
title_sort streamlined preparation of genomic dna in agarose plugs for pulsed-field gel electrophoresis
topic Protocol
url 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
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