Solution A(see below)............25.0 ml
Solution B(see below).............6.25 ml
KNO3 ...............................0.253 g
NaNO3 ..............................0.212 g
Purified agar (if necessary)......10.0 g
Distilled water..................969.0 ml
Combine ingredients, with the exception of Solution B, and autoclave at 121C for 15 min.
Prepare Solution B as instructed below, and aseptically add to basal medium.
Solution A:
4% MgSO4 . 7H2O ..............................500.0 ml
1.2% CaCl2 . 2H2O ............................500.0 ml
3.8% NaCl....................................500.0 ml
Microelements Stock Solution (see below).....500.0 ml
Microelements Stock Solution:
A & A FeEDTA Solution (see below)............160.0 ml
MnCl2 . 4H2O .................................360.0 mg
MoO3 ..........................................36.0 mg
ZnSO4 . 7H2O ..................................44.0 mg
CuSO4 . 5H2O ..................................15.8 mg
H3BO3 ........................................572.0 mg
NH4VO3 .........................................4.6 mg
CoCl2 . 6H2O ...................................8.0 mg
Distilled water.............................1090.0 ml
A & A FeEDTA Solution:
1. Dissolve 5.2 g KOH in 186 ml distilled water.
2. Add 20.4 g EDTA . 2H2O.
3. Dissolve 13.7 g FeSO4 . 7H2O in 364 ml distilled water.
4. Mix solutions 2 and 3.
5. Bubble Millipore-filtered air through solution until color changes. (may take minutes to hours.) Final pH of FeEDTA solution approximately 3.5.
Solution B:
K2HPO4 ......................28.0 g
Distilled water............500.0 ml
Autoclave at 121C for 15 minutes. Add aseptically to basal medium.