Desulfonatronovibrio Medium (DSMZ Medium 742)

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfonatronovibrio Medium
(DSMZ Medium 742)

培养基配方
Composition per liter:
NaHCO3 .......................................................................15.0g
Na2CO3 .......................................................................10.0g
NaCl..........................................................................10.0g
Na2SO4 ........................................................................3.0g
NH4Cl .........................................................................1.0g
Na2HPO4 .......................................................................0.2g
KCl........................................................................... 0.2g
Resazurin ....................................................................0.5mg
Yeast extract solution.......................................................10.0mL
Na-formate solution..........................................................10.0mL
Trace elements solution .....................................................10.0mL
Vitamin solution.............................................................10.0mL
Na2S·9H2O solution..........................................................10.0mL
pH 9.6 ± 0.2 at 25°C
Na-Formate Solution:
Composition per 10.0mL:
Na-formate.....................................................................5.0g
Preparation of Na-Formate Solution:  Add Na-formate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................1.5g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave under 100% N2 for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O......................................................................1.0g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to distilled/
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.
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. Filter sterilize.
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 100% N2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free atmosphere of 100% N2 . Add components, except vitamin
solution, yeast extract solution, Na-formate solution, trace elements so-
lution, and Na2S·9H2O solution, to distilled/deionized water and bring
volume to 950.0mL. Mix thoroughly. Sparge with 100% N2 . Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically and an-
aerobically add 10.0mL sterile vitamin solution, 10.0mL of sterile
yeast extract solution, 10.0mL sterile Na-formate solution, 10.0mL
sterile trace elements solution, and 10.0mL of sterile Na2S·9H2O solu-
tion. Mix thoroughly. Adjust pH to 9.6. Aseptically and anaerobically
distribute into sterile tubes or flasks.
Use:  For the cultivation of Desulfonatronovibrio hydrogenovorans.