Base

Heat with agitation to dissolve agar. Dispense 3 ml portions into 13 x 100 mm tubes. Autoclave 15 min at 121°C. Cool to 50°C; pH, 7.1.

Carbohydrate stock solution

Dissolve 10 g carbohydrate in 90 ml distilled water. Sterilize by filtration through 0.22 µm membrane. Add 0.3 ml stock solution to 2.7 ml base in tube. Mix gently and cool at room temperature.

Inoculate tubes in duplicate. Layer one tube with sterile mineral oil. Incubate 48 h at 35°C.

For use with halophilic Vibrio spp., add NaCl to a final concentration of 2-3%.

Hypertext updated by rim/cjm 2005-OCT-21