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山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Ancalomicrobium Medium
Composition per liter:
Glucose .......................................................................................0.25g
(NH4)2SO4 ...................................................................................0.25g
Na2HPO4 ...................................................................................0.071g
Hutner's basal salts solution.....................................................20.0mL
Vitamin solution.......................................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Hutner’s Basal Salts Solution:
Composition per liter:
MgSO4 ·7H2O..............................................................................29.7g
Nitrilotriacetic acid.....................................................................10.0g
CaCl2 ·2H2O...............................................................................3.335g
FeSO4 ·7H2O.............................................................................99.0mg
(NH4)6MoO7O24 ·4H2O ............................................................9.25mg
"Metals 44" ..............................................................................50.0mL
"Metals 44":
Composition per 100.0mL:
ZnSO4 ·7H2O.............................................................................1.095g
FeSO4 ·7H2O..................................................................................0.5g
Sodium EDTA.............................................................................0.25g
MnSO4 ·H2O .............................................................................0.154g
CuSO4 ·5H2O............................................................................39.2mg
Co(NO3)2 ·6H2O.......................................................................24.8mg
Na2B4O7 ·10H2O.......................................................................17.7mg
Preparation of Metals “44”: Add sodium EDTA to distilled/de-
ionized water and bring volume to 90.0mL. Mix thoroughly. Add a few
drops of concentrated H2SO4 to retard precipitation of heavy metal
ions. Add remaining components. Mix thoroughly. Bring volume to
100.0mL with distilled/deionized water.
Preparation of Hutner’s Basal Salts Solution:  Add nitrilotria-
cetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5
with KOH. Add remaining components. Add distilled/deionized water
to 1.0L. Adjust pH to 6.8. Filter sterilize.
Vitamin Solution:
Composition per liter:
Calcium  DL -pantothenate...........................................................5.0mg
Nicotinamide..............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Cyanocobalamin........................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Add components, except Hutner's basal
salts solution and vitamin solution, to distilled/deionized water and
bring volume to 970.0mL. Mix thoroughly. Adjust pH to 7.0. Auto-
clave for 15 min at 15 psi pressure–121°C. Aseptically add 20.0mL of
sterile Hutner's basal salts solution and 10.0mL of sterile vitamin solu-
tion. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Ancalomicrobium adetum.