NZY Agar (40 Plates)

Reagents

Protocol

- To make 2x NZY medium, dissolve 10 g NZ amine, 5 g yeast extract, and 5 g NaCl in 500 mL water.

- Adjust pH to 7.5 with NaOH, autoclave, and let reach room temperature.

- In a separate flask, mix 11 g agar and 500 mL water. Autoclave.

- To the autoclaved and not-cooled agar add the 500 mL  of 2× NZY medium.

-To minimize bubbles, pour the medium gently down the wall of the flask at an angle that prevents the medium from splashing down directly into the agar. 

- Mix the contents of the flask by gentle rotation, holding the flask at a shallow angle to promote mixing, but avoiding bubbles.

- Add antibiotics if using and pour the medium immediately onto plates adding at least enough to cover the bottom.

- Allow the plates to cool to room temperature. Dry the plates overnight at room temperature, or for a few hours at 37°C, upside down with the lids tilted to allow airflow. This should be done in a sterile laminar flow hood to avoid contamination of the plates.