Streamlining standard bacteriophage methods for higher throughput
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1-s2.0-S221501611830013X-main.pdf
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1.17 MB
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Adobe PDF
Checksum (MD5)
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Author(s)
Kauffman, Anne Kathryn Marie
Polz, Martin F
Date Issued
January 2018
Journal
MethodsX
Publisher
Elsevier BV
Citation
Kauffman, Kathryn M. and Martin F. Polz. “Streamlining Standard Bacteriophage Methods for Higher Throughput.” MethodsX 5 (2018): 159–172 © 2018 The Author(s)
Version
Final published version
Abstract
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 Keywords: Agar overlay; Agar layer; Top agar; Bottom agar; Plaque; Virus; Phage; Isolation; Cultivation; Purification
MIT Department
Massachusetts Institute of Technology. Center for Microbiome Informatics and Therapeutics
Massachusetts Institute of Technology. Department of Civil and Environmental Engineering
Terms of Use
Creative Commons Attribution 4.0 International License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/J.MEX.2018.01.007
