P22 Phage Lysates
Modified from Jeff Lawrence
- 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.
Last Update: Thursday, 19-Jun-2014 11:54:41 PDT
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