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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) .
Cowley Curent sheets in MHD