By Jan Michiels, Maarten Fauvart
This quantity provides a finished number of equipment which were instrumental to the present realizing of bacterial persisters. Chapters within the booklet hide issues starting from normal equipment for measuring persister degrees in Escherichia coli cultures, protocols for the decision of the persister subpopulation in Candida albicans, quantitative measurements of variety I and kind II persisters utilizing ScanLag, to in vitro and in vivo models for the research of the intracellular job of antibiotics. Written within the hugely winning Methods in Molecular Biology series layout, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, comfortably reproducible laboratory protocols, and pointers on troubleshooting and averting recognized pitfalls.
Authoritative and state of the art, Bacterial endurance: tools and Protocols brings jointly the main revered researchers in bacterial patience whose reports will stay important to figuring out this box for a few years to come.
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Additional info for Bacterial Persistence: Methods and Protocols
Acknowledgements This work was supported by the European Regional Development Fund through the Center of Excellence in Chemical Biology and Estonian Science Foundation grant 8822. References 1. Bigger JW (1944) Treatment of staphylococcal infections with penicillin by intermittent sterilization. Lancet 244(6320):497–500 2. Lewis K (2010) Persister cells. Annu Rev Microbiol 64:357–372 3. Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K (2004) Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 230 (1):13–18 4.
Grow on a shaker at 37 C overnight (see Note 8). 2. Make a 100Â dilution of the overnight culture into 20 mL of LB medium. Incubate on a shaker at 37 C. Sample over time to measure the optical density at λ ¼ 600 nm. 3. 8 mL of DMSO. Dispense in 100 μl aliquots in cluster tubes and store frozen at À80 C. DMSO stocks can be stored for up to 3 months. 2 Growing Overnight Cultures for Inoculum A standardized procedure for preparation of inocula makes experiments reproducible and is required for consistent results (see Note 9).
Slattery A, Victorsen AH, Brown A, Hillman K, Phillips GJ (2013) Isolation of highly persistent mutants of Salmonella enterica serovar Typhimurium reveals a new toxin-antitoxin module. J Bacteriol 195(4):647–657. doi:10. 01397-12 10. Lechner S, Lewis K, Bertram R (2012) Staphylococcus aureus persisters tolerant to bactericidal antibiotics. J Mol Microbiol Biotechnol 22(4):235–244. 1159/000342449 11. Leung V, Levesque CM (2012) A stressinducible quorum-sensing peptide mediates the formation of persister cells with noninherited multidrug tolerance.