【成份】
Trace metal solution:
1058 ALKALIPHILUS CROTONATOXIDANS MEDIUM
Yeast extract (BD-Difco) 2.0 g
Tryptone (BD-Difco) 2.0 g
Crotonic acid 0.86 g
KH2PO4 0.5 g
MgCl2·6H2O 0.33 g
NaCl 0.4 g
NH4Cl 0.4 g
CaCl2·2H2O 0.05 g
Trace metal solution (see below) 10.0 ml
Trace vitamins (见培养基0901) 10.0 ml
NaHCO3 3.5 g
L-Cysteine·HCL·H2O 0.5 g
Na2S·9H2O 0.5 g
Resazurin 1.0 mg
Dissolve ingredients
except crotonic acid
NaHCO3
L-cysteine·HCl·H2O and Na2S·9H2O
in 900 ml of distilled water and adjust pH to 7.0-7.5. Bring to a boil for 5-10 sec
then
cool down under a stream of N2-CO2 (80:20
v
) and add NaHCO3 to the medium. Dispense 0.9 volume of the medium into suitable culture vessels under a stream of N2-CO2 (80:20
v
) and seal with butyl rubber stoppers. After autoclaving
stand the medium overnight. Add 0.1 volume of a filter-sterilized crotonic acid solution (0.86 g/100 ml
pH 7.0) to the medium. Autoclave cysteine·HCl and Na2S·9H2O as 5% solutions
respectively
under a N2 atmosphere. Prior to inoculation
aseptically and anaerobically add the solutions of crotonic acid
L-cysteine·HCl·H2O and Na2S·9H2O to the medium.
MnSO4·xH2O 1.0 g
Fe(NH4)2(SO4)2·6H2O 0.8 g
CoCl2·6H2O 0.2 g
ZnSO4·7H2O 0.2 g
CuCl2·2H2O 0.02 g
NiCl2·6H2O 0.02 g
Na2MoO4·2H2O 0.02 g
Na2SeO4 0.02 g
Na2WO4 0.02 g
Nitrilotriacetic acid 2.0 g
Distilled water 1.0 L
Adjust pH to 6.0 with KOH.