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Streamlining standard bacteriophage methods for higher throughput

A universal tool in the culture-based study of bacterial viruses (bacteriophages, or phages) is the agar overlay, which is used in the isolation of new viruses, and in their quantification and purification. Here, simple optimizations that increase efficiency and throughput in agar overlay based isol...

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Detalles Bibliográficos
Autores principales: Kauffman, Kathryn M., Polz, Martin F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6318102/
https://www.ncbi.nlm.nih.gov/pubmed/30622914
http://dx.doi.org/10.1016/j.mex.2018.01.007
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author Kauffman, Kathryn M.
Polz, Martin F.
author_facet Kauffman, Kathryn M.
Polz, Martin F.
author_sort Kauffman, Kathryn M.
collection PubMed
description A universal tool in the culture-based study of bacterial viruses (bacteriophages, or phages) is the agar overlay, which is used in the isolation of new viruses, and in their quantification and purification. Here, simple optimizations that increase efficiency and throughput in agar overlay based isolation and cultivation of virus-host systems are presented. The agar overlay is streamlined to minimize steps and materials. Serial purification of viruses from viral colonies (plaques) is optimized to eliminate steps by combining purification by serial re-streaking with the optimized agar overlay approach. Finally, recommendations are made for efficient archival and storage of virus plaques. In sum, this work presents: • Tube-free Agar Overlays: rapid plaque assays with fewer steps and materials; • Molten Streaking for Singles: rapid tube-free serial purification of viruses; • Archiving Plaques: saving virus purification for later.
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spelling pubmed-63181022019-01-08 Streamlining standard bacteriophage methods for higher throughput Kauffman, Kathryn M. Polz, Martin F. MethodsX Immunology and Microbiology A universal tool in the culture-based study of bacterial viruses (bacteriophages, or phages) is the agar overlay, which is used in the isolation of new viruses, and in their quantification and purification. Here, simple optimizations that increase efficiency and throughput in agar overlay based isolation and cultivation of virus-host systems are presented. The agar overlay is streamlined to minimize steps and materials. Serial purification of viruses from viral colonies (plaques) is optimized to eliminate steps by combining purification by serial re-streaking with the optimized agar overlay approach. Finally, recommendations are made for efficient archival and storage of virus plaques. In sum, this work presents: • Tube-free Agar Overlays: rapid plaque assays with fewer steps and materials; • Molten Streaking for Singles: rapid tube-free serial purification of viruses; • Archiving Plaques: saving virus purification for later. Elsevier 2018-01-31 /pmc/articles/PMC6318102/ /pubmed/30622914 http://dx.doi.org/10.1016/j.mex.2018.01.007 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Immunology and Microbiology
Kauffman, Kathryn M.
Polz, Martin F.
Streamlining standard bacteriophage methods for higher throughput
title Streamlining standard bacteriophage methods for higher throughput
title_full Streamlining standard bacteriophage methods for higher throughput
title_fullStr Streamlining standard bacteriophage methods for higher throughput
title_full_unstemmed Streamlining standard bacteriophage methods for higher throughput
title_short Streamlining standard bacteriophage methods for higher throughput
title_sort streamlining standard bacteriophage methods for higher throughput
topic Immunology and Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6318102/
https://www.ncbi.nlm.nih.gov/pubmed/30622914
http://dx.doi.org/10.1016/j.mex.2018.01.007
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