3-Color Infection Assay

This protocol is used to check for phage E. coli receptor usage using three one-receptor strains containing only the LamB, OmpF, or OmpC receptor each. The phage are plated on a plate containing 1:1:1 ratio of the three strains, each containing a different fluoroscent reporter, and then imaged using a fluoroscent microscope setup.

Strains:

• 3-color strains
  • E. coli \(\Delta\)LamB \(\Delta\)OmpF w/ mClover3 (eOC)
  • E. coli \(\Delta\)OmpF \(\Delta\)OmpC w/ mTurquoise2 (eLB)
  • E. coli \(\Delta\)OmpC \(\Delta\)LamB w/ mScarlet-I (eOF)

Reagents: LB Media, 0.3% NZCYM Agar, 1000x Kanamycin stock, phage sample.

Time required: 2 days (excluding day 0)