Dissolve ingredients (except fumarate and NaHCO3), bring to a
boil, cool down to room temperature while gasing the medium with 80% N2
+ 20% CO2, and add the bicarbonat. Equilibrate the medium with
gas to reach a pH of 6.8. Dispense the medium in anaerobe vials under N2
+ CO2. After autoclaving add fumarate from an anaerobic filter
sterilized stock solution (e.g. 16% w/v Na2-fumarate; add 0.5
ml/10 ml medium). Check medium pH which should be around 6.8.
Trace element solution
Distilled water
1000.00
ml
Na2SeO3 x 5 H2O
0.30
mg
NiCl2 x 6 H2O
0.03
g
Na2MoO4 x 2 H2O
0.01
g
H3BO3
0.01
g
KAl(SO4)2 x 12 H2O
0.02
g
CuSO4 x 5 H2O
0.01
g
ZnSO4 x 7 H2O
0.18
g
CaCl2 x 2 H2O
0.10
g
CoSO4 x 7 H2O
0.18
g
FeSO4 x 7 H2O
0.10
g
NaCl
1.00
g
MnSO4 x H2O
0.50
g
MgSO4 x 7 H2O
3.00
g
Nitriloacetic acid
1.50
g
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then
add minerals. Final pH 7.0 (with KOH).
