【成份】
1055
sodium acetate 0.82 g
CaCl2·2H2O 0.147 g
NH4Cl 0.535 g
NaH2PO4 0.136 g
Na2SO4 0.142 g
Trace element solution 10.0 ml
Vitamins solution(见培养基0901) 10.0 ml
Se/W solution(见培养基0975) 1.0 ml
Distilled water 1.0 L
Mix components thoroughly and autoclave under a N2-CO2 (80:20
v
) gas mixture. After cooling
add aseptically and anaerobically the following solutions to the medium:
8% NaHCO3 solution (filter-sterilized) 30.0 ml
100 mM Fe NTA solution (see below) 50.0 ml
Adjust pH to 7.0 - 7.2
100 mM Fe NTA solution:
NaHCO3 1.64 g
Nitrilotriacetic acid trisodium salt monohydrate 2.75 g
FeCl3·6H2O 2.7 g
Firstly
dissolve NaHCO3 in 80 ml distilled water. Then
add NTA and FeCl3·6H2O to the solution and bring volume to 100 ml with distilled water. Adjust pH to 7.0 with NaOH. Filter-sterilize and store under a N2 atmosphere
Trace element solution:
Nitrilotriacetic acid 12.8 g
FeCl3·6H2O 1.35 g
MnCl2·4H2O 0.1 g
CoCl2·6H2O 0.024 g
CaCl2·2H2O 0.1 g
ZnCl2 0.1 g
CuCl2·2H2O 0.025 g
H3BO3 0.01 g
NaCl 1.0 g
NiCl2·6H2O 0.12 g
Na2SeO3·5H2O 0.026 g
Distilled water 1.0 L
Dissolve nitrilotriacetic acid first in 200 ml of distilled water
and adjust pH to 6.5 with KOH. Then add the remaining salts and adjust volume to 1.0 L. Readjust pH to 7.0 with KOH.