![]() ![]() cereus VKM B-370 culture (OD = 1.2) and 445 µL of LB broth with 10 mM CaCl 2 and 10 mM MgCl 2 in a well of a 48-well microplate. In order to propagate the phage, 50 µL of each phage extract was mixed with 5 µL of a B. The obtained phage extracts were briefly centrifuged at 3000 g. Separate plaques were mixed with 750 μL of SM+ buffer (50 mM Tris-HCl, pH 8.0 100 mM NaCl 1 mM MgSO 4 0.1% gelatin 10 mM CaCl 2 10 mM MgCl 2) and 75 μL of chloroform in a 1.5-mL Eppendorf tube followed by incubation overnight at 4 ☌ for phage extraction. cereus VKM B-370 using the double agar layer technique, and the plate was incubated at 28 ☌ overnight. The supernatant was titrated with serial dilutions on the lawn of the sensitive strain B. The cell debris was removed by centrifugation at 3000 g for 12 min at 4 ☌, and 3 μL of chloroform was added to the resulting supernatant. The culture was incubated at 28 ☌ until optical density decreased. Then, 5 µL of mitomycin C (50 μg/mL) was added to a final concentration of 0.5 μg/mL. The microplate was incubated with shaking at 28 ☌ in a FilterMax F5 microplate reader (Molecular Devices), with OD 595 measured every 10 min. Briefly, the bacterial strain was grown in a well of a 48-well microplate, in 495 µL of LB broth with 10 mM CaCl 2 and 10 mM MgCl 2, until the optical density reached 0.2 at 595 nm. cereus VKM B-13 was performed using mitomycin C. The induction of phage B13 from the lysogenic strain B.
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