Ferroglobus placidus Medium (DSMZ Medium 730)

[所属分类:培养基配方] [发布时间:2020-7-15] [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

Ferroglobus placidus Medium
(DSMZ Medium 730)

培养基配方
Composition per 1020.0mL:
NaCl.......................................................................18.0g
NaHCO3 ....................................................................10.0g
MgCl2 ·6H2O ...............................................................4.3g
KNO3 .......................................................................1.0g
KCl........................................................................0.34g
NH4Cl .....................................................................0.24g
CaCl2 ·2H2O ..............................................................0.14g
K2HPO4 ·3H2O .............................................................0.14g
Resazurin .................................................................0.5mg
Trace elements solution...................................................10.0mL
Vitamin solution..........................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Na-pyruvate solution .....................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Na-pyruvate Solution:
Composition per 10.0mL:
Na-pyruvate ................................................................1.0g
Preparation of Na-pyruvate Solution: Add Na-pyruvate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Trace Elements Solution:
Composition per liter:
MgSO4 ·7H2O.................................................................3.0g
Nitrilotriacetic acid........................................................1.5g
NaCl.........................................................................1.0g
MnSO4 ·2H2O.................................................................0.5g
CoSO4 ·7H2O................................................................0.18g
ZnSO4 ·7H2O................................................................0.18g
CaCl2 ·2H2O ...............................................................0.1g
FeSO4 ·7H2O................................................................0.1g
NiCl2 ·6H2O..............................................................0.025g
KAl(SO4)2 ·12H2O..........................................................0.02g
H3BO3 .....................................................................0.01g
Na2MoO4 ·4H2O.............................................................0.01g
CuSO4 ·5H2O...............................................................0.01g
Na2SeO3 ·5H2O.............................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl............................................................10.0mg
Thiamine-HCl·2H2O.........................................................5.0mg
Riboflavin ................................................................5.0mg
Nicotinic acid.............................................................5.0mg
D -Ca-pantothenate.........................................................5.0mg
p-Aminobenzoic acid........................................................5.0mg
Lipoic acid ...............................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Vitamin B 12 ..............................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O..................................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Sparge with N2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Preparation of Medium: Add components, except Na2S·9H 2 O so-
lution, vitamin solution, and Na-pyruvate solution, to distilled/deion-
ized water and bring volume to 990.0mL. Mix thoroughly. Flush
medium with N2 for 20 min. Adjust medium pH to 7.0 with 4N H2SO4 .
Add 10.0mL of Na2S·9H2O solution. Mix thoroughly. Readjust the
medium pH to 7.0 with H2SO4 , while flushing the gas phase only with
N2 . Dispens 10mL volumes into 100mL serum bottles with rubber
stoppers under N2 . Replace gas phase by 80% H2 + 20% CO2 gas mix-
ture and finally pressurize the bottles to 2 bar gas overpressure. Auto-
clave for 15 min at 15 psi pressure–121°C. Aseptically inject via syringe
0.1mL vitamin solution and 0.1mL Na-pyruvate solution into each tube
containing 10.0mL of the autoclaved medium.
Use:  For the cultivation of Ferroglobus placidus.