P22 Phage Lysates

Protocol

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Grow host strain overnight in LB.

Dilute bacteriophage P22 stock to 10⁸ pfu/mL (about a 10^-2 dilution)

In a 10 mL culture tube, combine 100 µL of diluted phage with 4.7 mL of Transducing Broth:

1xLB medium (100 mL LB)

1xE medium (2 mL 50X E medium)

0.2% Glucose (400 µL 50% Glucose)

[At this point, the diluted phage can be stored indefinitely at 4°C]

Add 1 mL cells to 4 mL phage broth and incubate overnight at 37°C with shaking.

Add 100 µL chloroform and incubate 5 min at 37°C with shaking.

Spin in tabletop centrifuge at high speed for 20 min at 4°C.

Transfer supernatant to a sterile screwcap tube.

Add 100 µL chloroform and store at 4°C.