Medium data

Search for medium no. = [1398].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.


Yeast extract (BD-Difco)  1.00    g
Glycyl-glycine  1.00    g
Na2HPO4·2H2O  0.10    g
MgSO4·7H2O  0.10    g
KNO3  0.10    g
NaNO3  0.50    g
NaCl  0.10    g
CaCl2·2H2O  0.05    g
Fe(III) citrate solution (0.1 g in 100 ml)  5.00    ml
SL-6 trace element solution (see Medium No. 167)  1.00    ml
Distilled water  1050.00    ml
Adjust pH to 8.2. 
Boil the medium under a stream of nitrogen gas for a few minutes and distribute 90 ml medium into 100 ml screw-capped bottles. Bubble each bottle with nitrogen gas and close immediately with a rubber septum and screw tight. Autoclave at 121C for 15 min. After autoclaving inject 1.0 ml of neutralized sulfide solution (0.03% final concentration) and 0.1 ml sterile vitamin solution CA. 
Neutralized sulfide solution:
Distilled water  100.00    ml
Na2S·9H2O  3.00   
The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed, and autoclaved for 15 min. at 121C. After cooling to room temperature, the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle.  Appearance of a yellow color indicates the drop of pH to about 8. The solution should be stirred continuously to avoid precipitation of elemental sulfur. The final solution should be clear and is yellow in color.   
Vitamin solution CA: 
Distilled water  100.0     ml
Nicotinic acid  100.0     mg
Thiamine-HCl·2H2O  100.0     mg
Biotin  5.0     mg
p-Aminobenzoic acid  50.0     mg
Vitamin B12  1.0     mg
Ca-pantothenate  50.0     mg
Pyridoxine-HCl  50.0     mg
Folic acid  50.0     mg
Na3-EDTA  200.0     mg
Adjust pH to 7.5. 
The solution is filter-sterilized. 

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