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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Journal of Biological Methods
2018
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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. |
format | Online Article Text |
id | pubmed-6706164 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Journal of Biological Methods |
record_format | MEDLINE/PubMed |
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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