P22 Phage Lysates

Modified from Jeff Lawrence
Protocol

==========

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
This page has been viewed 392 times.
Eric Kofoid eckofoid at ucdavis.edu