Plaque assay - glasgowlab/home GitHub Wiki

Important: think about what cells you use. If your sample or cells require antibiotics, use the appropriate plate.

Making top agar = 50% LB and 50% LB agar. Lb agar should be melted before mixing.

Spotting:

  1. Mix 150 uL of overnight culture with 3 mL hot (melted) top agar. Vortex very gently just to make sure everything is mixed.
  2. Pour onto LB plate and tilt the plate to spread everything out (do this quickly before the top agar cools and solidifies
  3. Wait 10 minutes for everything to dry
  4. Multichannel pipette 2 uL of your serial dilutions (the thing your are testing for phage)
  5. Wait 15 minutes for everything to dry (if you jostle the plate, your spots will spread and run into each other!! This is why the waiting is important!!)
  6. Place in 37C incubator.