20.109(F07): Growth of phage materials

Protocols

In advance of this lab, a bacterial host (namely XL1-blue) was infected with the modified M13 phage called "3-12." Though the precise details of the 3-12 modification are still unpublished, it was isolated from a screen for changes in the M13 p8 that enable the phage to bind a metal (Iridium). Today you will harvest the phage from the supernatant of the infected bacterial culture and then titer it.

Part 1: Phage purification

1. Spin 2 eppendorf tubes with infected cells in a room temperature microfuge, 1 minute at full speed.
2. Remove the supernatant to clean fresh eppendorf tubes.
3. Use your P1000 to measure the volume, then add a 1/6th volume of a 20% PEG-8000/2.5M NaCl solution.
4. Invert to mix then incubate on ice 60 minutes.
5. Spin in a room temperature microfuge, 15 minutes at full speed.

Part 2: Titering phage

DONE!