1. Dry NZY plates for 30-60 minutes at 37 °C or age at room temperature
for a couple of days.
2. Melt NZY top agarose (not agar) and place in 45-50 °C water bath. 3 ml top agarose is needed for one 100 mm plate.
3. Make a serial dilution (1:100) of the phage stock in
10 mM Tris-HCl pH7.5, 10 mM MgCl2, 50 mM NaCl.
4. Mix 100 µl of infection-ready host cells to 100 µl of diluted phage for one 100 mm plate.
5. Incubate at 37 °C for 15-20 minutes.
6. (optional) For libraries with blue/white selection, add X-Gal and IPTG to the top agarose.
7. Add 3 ml NZY top agarose (45-50 °C) to each 200 µl of infected cells.
8. Mix and quickly pour onto the center of a 100 mm NZY plate. Use 6.25 ml of NZY top agarose per 150 mm plate.
9. Gently swirl to ensure even distribution.
10. Allow NZY top agarose to set .
11. Invert plates and incubate overnight at 37 °C.
12. Count the number of plaques.