Base Medium

Biochemicals (see below)

Neutral red solution, 0.2% Dissolve 0.2 g neutral red in 10 ml EtOH in a 100 ml volumetric and bring to line with d. water.

Mix base medium ingredients, then boil. Divide into four 250 ml portions. Add 2.5 ml neutral red to 3 of the 4 portions. Add glycine, NaCl and cysteine-HCl to the 3 portions containing neutral red. Add potassium nitrate in the portion without neutral red. Adjust the pH of each portion to 7.4 ± 0.2. Dispense 10 ml per 16 X 125 mm screw-cap tube. Autoclave 121°C, 15 min.

1. Potassium nitrate (for Final Concentration of 1% (w/v)). Add 2.5 g to 250 ml (10 g/liter) semi-solid mixture without neutral red.
2. Glycine (for Final Concentration of 1% (w/v)). Add 2.5 g to 250 ml (10 g/liter) semi-solid mixture with neutral red.
3. NaCl (for Final Concentration of 3.5% (w/v)). Add 7.5 g to 250 ml (30 g/liter) semi-solid mixture with neutral red.
4. Cysteine-HCl (for Final Concentration of 0.02% (w/v)). Add 0.05 g to 250 ml (0.2 g/liter) semi-solid mixture with neutral red.

Hypertext updated by rim/kwg/cjm 2005-OCT-21