Bacteriophages: Methods and Protocols, Volume 1: Isolation, - download pdf or read online

By K. Eric Wommack, Kurt E. Williamson, Rebekah R. Helton, Shellie R. Bench (auth.), Martha R.J. Clokie, Andrew M. Kropinski (eds.)

Ranging from the evolution of pathogenicity to oceanic carbon biking, the various and sundry roles that bacteriophages play in microbial ecology and evolution have encouraged elevated curiosity in the clinical neighborhood. Bacteriophages: tools and Protocols pulls jointly the significant physique of information and services from most sensible foreign bacteriophage researchers to supply either classical and cutting-edge molecular recommendations. With its well-organized modular layout, Volume 1: Isolation, Characterization, and Interactions examines a large number of subject matters, together with the isolation of phages, morphological and molecular characterization, and interplay with micro organism. Written within the hugely profitable Methods in Molecular Biology™ sequence layout, chapters include short introductions to the topic, lists of the required fabrics and reagents, easily reproducible laboratory protocols, and a Notes part which information pointers on troubleshooting and keeping off recognized pitfalls.

Thorough and state-of-the-art, Bacteriophages: tools and Protocols is a worthy reference for skilled bacteriophage researchers in addition to an simply obtainable advent for novices to the subject.

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Individual well-separated plaques will be visible after overnight incubation. 4 Notes 1. It is recommended that careful attention should be paid to collecting the samples in such a way as not to contaminate the outside of the storage container or the person collecting 20 Van Twest and Kropinski 2. 3. 4. 5. 6. 7. 8. 9. the samples. Please note that raw sewage will most definitely contain human pathogens. For the bacteria listed above, Difco Luria broth or Tryptic Soy Broth work well. de/). Many phages require 1–10 mM divalent ions such as Ca2+ or Mg2+ for attachment or intracellular growth (18, 19, 20).

1999). Evolutionary relationships among diverse bacteriophages and prophages: all the world’s a phage. Proc. Natl. Acad. Sci. USA 96, 2192–2197. 7. Moreira, D. (2000). Multiple independent horizontal transfers of informational genes from bacteria to plasmids and phages: implications for the origin of bacterial replication machinery. Mol. Microbiol. 35: 1–5. 8. F. (2005). Bacteriophages and bacterial virulence, in Bacteriophages: Biology and Applications, (Kutter E. ), CRP Press, FL, pp. 223–265.

Irradiate with constant swirling (120 rpm) for 20–30 s. Petri dish should be placed 16 cm from the germicidal short wave lamp. 28 Raya and Hébert 7. Transfer cells to a screw-capped tube containing 5 ml double strength (MRS or M17) broth. 8. Incubate at 30◦ C (lactococci) or 37◦ C (lactobacilli) in a constant temperature water bath. Protect cells from light exposure. 9. 05M MgSO4 as control zero of absorbance). Plot time (h) versus absorbance (600 nm). 10. Centrifuge lysate at 3000 × g for 12 min at 4◦ C.

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