By Alain Filloux, Juan-Luis Ramos
In Pseudomonas aeruginosa, specialist researchers within the box aspect some of the tools that are now well-known to check this attention-grabbing microorganism. Chapters comprise microbiological how to high-throughput molecular strategies which were built during the last decade. Written within the hugely winning Methods in Molecular Biology sequence structure, chapters contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, quite simply reproducible laboratory protocols and key tips about troubleshooting and heading off identified pitfalls.
Authoritative and functional, Pseudomonas aeruginosa aids within the carrying on with learn of recent and leading edge findings.
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Additional info for Pseudomonas Methods and Protocols
If agarose blocks do not harden in the mold, the agarose concentration is too low.
10. Transfer the DNA–cell mix in an electroporation cuvette (see Note 7). 11. Place the electroporation cuvette containing the DNA–cell mix in the Gene Pulser. Set the parameters as described in Subheading 2 and apply the electric shock (see Note 8). 14 Frédéric Cadoret et al. 12. Transfer the mix in 12 ml falcon tube containing 2 ml of SOC glucose medium. 13. Cultivate the cells for 2 h at 37 °C under shaking conditions (see Note 9). 14. Display 200 μl of the bacterial culture on selective agar plates (see Note 10) and incubate at 37 °C for at least 48 h.
Here, we describe the methods used to isolate, concentrate, purify, and produce bacteriophages specific for Pseudomonas species. Key words Bacteriophage, Enrichment, Isolation, Titration, Production 1 Introduction Bacteriophages are bacterial viruses hypothesized to exist wherever their hosts thrive. The vast majority of known bacteriophages, including those targeting pseudomonads, belong to the Caudovirales order comprising three families of dsDNA bacteriophages differing in their tail characteristics: long and non-contractile (Siphoviridae), long and contractile (Myoviridae), and short and non-contractile (Podoviridae) .
Pseudomonas Methods and Protocols by Alain Filloux, Juan-Luis Ramos