Archaeoglobus veneficus Medium (DSMZ Medium 796)

[所属分类:培养基配方] [发布时间:2019-11-7] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Archaeoglobus veneficus Medium
(DSMZ Medium 796)
Composition per liter:
NaCl............................................................................................18.0g
MgCl2 ·6H2O ...............................................................................7.15g
NaHCO3 ........................................................................................5.0g
KCl..............................................................................................0.33g
NH4Cl .........................................................................................0.25g
K2HPO4 ·3H2O ............................................................................0.18g
CaCl2 ·2H2O ................................................................................0.14g
Fe(NH4)2(SO4)2 ·6H2O...............................................................2.0mg
Resazurin...................................................................................0.5mg
Na2SO3  solution.......................................................................20.0mL
Na2S·9H2O solution.................................................................10.0mL
Na-acetate solution....................................................................4.0mL
Trace elements solution.............................................................1.0mL
pH 1.0 ± 0.2 at 25°C
Na2S·9H2O Solution:
Composition per 20.0mL:
Na2S·9H2O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 20.0mL. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Na2SO 3 Solution:
Composition per 10.0mL:
Na2SO3 ..........................................................................................0.5g
Preparation of Na2SO3 Solution : Add Na2SO3 to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Filter sterilize.
Na-acetate Solution:
Composition per 10.0mL:
Na-acetate.....................................................................................2.5g
Preparation of Na-acetate Solution : Add Na-acetate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
Trace Elements Solution:
Composition per liter:
NaCl..............................................................................................5.0g
MnCl2 ·4H2O .................................................................................2.9g
(NH4)2Ni(SO4) 2 ............................................................................1.0g
FeSO4·7H2O..................................................................................0.5g
CoCl2·6H2O..................................................................................0.5g
CaCl2·2H2O ..................................................................................0.5g
ZnSO4·7H2 ....................................................................................0.5g
CuSO4 ·5H2O...............................................................................0.05g
H3BO3 .........................................................................................0.05g
KAl(SO4)2 ·12H2O.......................................................................0.05g
Na2MoO4 ·4H2O ..........................................................................0.05g
Na2WO4 ·2H2O............................................................................0.05g
Na2SeO3 ·5H2O............................................................................0.05g
Preparation of Trace Elements olution:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 .
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2  gas mixture. Add components, except Na-acetate solu-
tion, Na 2 S·9H 2 O solution, and NaHCO 3 , to 986.0mL distilled/deion-
ized water. Mix thoroughly. Gently heat and bring to boiling. Boil for
5 min. Cool to 25°C while sparging with 80% N 2 + 20% CO 2 . Add the
solid NaHCO 3 . Equilibrate with the 80% N 2 + 20% CO 2 gas. Adjust
pH to 7.0 with HCl. Add 10.0mL Na 2 S·9H 2 O solution. Distribute into
serum bottles under 80% N 2 + 20% CO 2 gas; 25.0mL medium in
100mL serum bottles. Adjust pH to 1.0. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 25°C. Aseptically and anaerobically inject
0.1mL sterile Na-acetate solution and 0.5mL Na 2 SO 3  solution per
25.0mL medium. Mix thoroughly. After inoculation pressurize to 1 bar
atmosphere with 80% H 2 + 20% CO 2 gas.
Use:  For the cultivation of Archaeoglobus veneficus.