Ferrous sulfate solution added prior to inoculation to the final concentration of 2 % (w/v)
Dissolve ingredients, adjust pH to 2.5 with H2SO4
and autoclave. Prior to inoculation add ferrous sulfate from a freshly
prepared 40 % (w/v) stock solution.
Ferrous sulfate solution
Prepare a 40 % (w/v) stock solution of FeSO4 x 7 H2O
in 0.1 N H2SO4. Sterilize by filtration and store
under 100 % N2 gas atmosphere.
Trace element solution
Nitrilotriacetic acid
1.50
g
MgSO4 x 7 H2O
3.00
g
MnSO4 x H2O
0.50
g
NaCl
1.00
g
FeSO4 x 7 H2O
0.10
g
CoSO4 x 7 H2O
0.18
g
CaCl2 x 2 H2O
0.10
g
ZnSO4 x 7 H2O
0.18
g
CuSO4 x 5 H2O
0.01
g
KAl(SO4)2 x 12 H2O
0.02
g
H3BO3
0.01
g
Na2MoO4 x 2 H2O
0.01
g
NiCl2 x 6 H2O
0.03
g
Na2SeO3 x 5 H2O
0.30
mg
Na2WO4 x 2 H2O
0.40
mg
Distilled water
1000.00
ml
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then
add minerals. Adjust final pH to 7.0 with KOH.