Isolation of Phage DNA

To separate the phage, the phage inducing lysate was centrifuged at 10,000 ×g for 15 min at 4°C and the supernatant was filtered through a 0.22 μm low protein binding filter (Millipore, United States). Contaminating nucleic acids in the supernatant were digested with pancreatic DNase I and RNase A, and each was added to obtain a final concentration of 10 μg/mL (Sigma-Aldrich Canada Ltd., Oakville, ON, United States) and incubated for 15 min at 37°C. DNA isolation was then performed according to Molecular Cloning: A Laboratory Manual, Third Edition (Sambrook J., Russell D.W. Molecular Cloning: A Laboratory Manual. 3rd ed. Volme 1 Cold Spring Harbor Press; Cold Spring Harbor, NY, United States: 2001), with minor modifications. For phage DNA purification the suspension was extracted twice with an equal volume of phenol-chloroform, once with chloroform and precipitated with ethanol. The pellets were washed in 70% ethanol, vacuum dried, and resuspended in 20 ml distilled water. PCR verification was performed on 1 ml of the phage DNA preparation.