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Plaque assay
Anum Azam Glasgow edited this page Jul 8, 2024
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1 revision
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:
- Mix 150 uL of overnight culture with 3 mL hot (melted) top agar. Vortex very gently just to make sure everything is mixed.
- Pour onto LB plate and tilt the plate to spread everything out (do this quickly before the top agar cools and solidifies
- Wait 10 minutes for everything to dry
- Multichannel pipette 2 uL of your serial dilutions (the thing your are testing for phage)
- 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!!)
- Place in 37C incubator.
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