1. Inoculate a single colony of host strain XL1-Blue
(MRF) into 5 ml LB containing 12.5 mg/ml tetracycline.
2. Incubate at 37°C for 1 hour with shaking.
3. Add 1 m helper phage (VCSM13 or RE704) or
pick from single plaques.
4. Incubate at 37 °C for 1 hour.
5. Add kanamycin to a final concentration of
6. Grow at 37 °C for 8 hours to overnight with shaking.
7. Pellet cell debris by centrifugation.
8. Aliquot the supernatant, titer and store at -20°C.