Luria-Bertani is the most preferred media to be used in phage research. This protocol describes how to make 1 L of Luria-Bertani agar (also known as LB agar) for the culturing of bacteria on plates.
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| ready-made Luria Bertani Agar plates |
Procedures/Steps
- Add 25g LB broth, Miller, and 15g agar (or 40g LB agar, Miller) per liter of water.
- Mix well by inverting the bottle several times until powder is dissolved.
- Mix well by inverting the bottle several times until powder is dissolved.
- Sterilize by autoclaving for 20 min at 15 psi (1.05 kg/cm2) on a liquid cycle.
- Following autoclave (while media are still liquid but cool enough to safely handle bottle) pour LB agar from the bottle into plates within a laminar flow hood. Allow ~50 mL per plate. The LB agar bottle must only be opened and the plates poured in a laminar flow hood to prevent contamination.
- Leave plates in a laminar hood until agar sets (~30 mins).
- When agar has set, replace the lid, invert plates, and store in an air-tight bag at 4 °C.
- Plates should be removed from 4 °C at least 1 h before use and placed in 30 °C incubators. To prevent condensation from the lid dropping onto the agar surface, wipe lids dry with Kim wipe and store plates inverted.
Reminder: only remove lid from LB agar plate while in a laminar flow hood to prevent contamination.
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