培養基配方
MgCl2·6H2O 0.2g
CaCl2·2H2O 0.1g
NaNO3 1.7g
NH4Cl 0.1g
KH2PO4 0.1g
KCl 0.1g
Trace element mixture* 1ml
Selenite-tungstate solution** 1ml
Bicarbonate solution*** 30ml
Vitamin solution**** 1ml
Thiamine solution***** 1ml
Vitamin B12 solution****** 1ml
Thiosulfate solution******* 10ml
Distilled water 1L
pH 7.2-7.4
Mix ingredients except for bicarbonate, vitamin, thiamine, cyanocobalamin and thiosulfate solutions. Dispense the medium into suitable culture vessels under a stream of N2/CO2/O2 (70/20/10), seal with butyl rubber stoppers, and autoclave at 121°C for20 min. After autoclaving, aseptically add other ingredients. Final pH is 7.2-7.4.
*Trace element mixture
HCl (25%; 7.7 M) 12.5ml
FeSO4·7H2O 2100mg
H3BO3 30mg
MnCl2·4H2O 100mg
CoCl2·6H2O 190mg
NiCl2·6H2O 24mg
CuCl2·2H2O 2mg
ZnSO4·7H2O 144mg
Na2MoO4·2H2O 36mg
Distilled water 987ml
Sterilized by autoclaving.
**Selenite-tungstate solution
NaOH 0.4g
Na2SeO3·5H2O 6mg
Na2WO4·2H2O 8mg
Distilled water 1L
Sterilized by autoclaving.
***Bicarbonate solution (1.0 M)
NaHCO3 8.4g
Distilled water 100ml
Saturated with CO2 gas. Tightly sealed in bottles filled with CO2 gas and then sterilized by autoclaving.
Vitamine solution
p-Aminobenzoic acid 4mg
Biotin 1mg
Nicotinic acid 10mg
Ca-pantothenate 5mg
Pyridoxine-HCl 15mg
Sodium phosphate buffer (10 mM; pH7.1) 100ml
Sterilized by filtration.
Thiamine solution
Thiamine-HCl 10mg
Sodium phosphate buffer (25 mM; pH 3.4) 100ml
Sterilized by filtration.
Vitamin B12 solution
Cyanocobalamin 5mg
Distilled water 100ml
Sterilized by filtration.
Thiosulfate solution (1.0 M)
Na2S2O3·5H2O 24.8g
Distilled water 100ml
Sterilized by filtration and stored under N2 gas.
