Oceanithermus profundus Medium (DSMZ Medium 975)

[所属分类:培养基配方] [发布时间:2021-6-25] [发布人:网站管理员2] [阅读次数:] [返回]

Oceanithermus profundus Medium
(DSMZ Medium 975)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

NaCl............................................................................................30.0g
HEPES........................................................................................2.38g
NH 4 Cl .........................................................................................0.33g
KCl..............................................................................................0.33g
Potassium nitrate solution........................................................10.0mL
Calcium chloride solution........................................................10.0mL
Magnesium chloride solution...................................................10.0mL
Yeast extract solution...............................................................10.0mL
Sucrose solution.......................................................................10.0mL
Tryptone solution.....................................................................10.0mL
Vitamin solution.........................................................................1.0mL
Trace elements solution .............................................................1.0mL
pH 7.0–7.5 at 25°C
Potassium Nitrate Solution:
Composition per 10.0mL:
KNO 3 ..........................................................................................0.33g
Preparation of Potassium Nitrate Solution: Add KNO 3 to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper-
ature.
Magnesium Chloride Solution:
Composition per 10.0mL:
MgCl 2 ·6H 2 O................................................................................0.33g
Preparation of Magnesium Chloride Solution: Add 0.33g of
MgCl 2 ·6H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Calcium Chloride Solution:
Composition per 10.0mL:
CaCl 2 ·2H 2 O.................................................................................0.33g
Preparation of Calcium Chloride Solution: Add CaCl 2 ·2H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................................0.2g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Sucrose Solution:
Composition per 10.0mL:
Sucrose..........................................................................................2.0g
Preparation of Sucrose Solution: Add sucrose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for
15 min at 15 psi pressure–121°C. Cool to room temperature.
Tryptone Solution:
Composition per 10.0mL:
Tryptone........................................................................................1.0g
Preparation of Tryptone Solution: Add tryptone to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................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. Sparge with 100% N 2 . Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................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% H 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 100%
N 2  gas atmosphere. Add components, except yeast extract solution, su-
crose solution, calcium chloride solution, magnesium chloride solu-
tion, potassium nitrate solution, tryptone solution, vitamin solution,
and trace elements solution, to distilled/deionized water and bring vol-
ume to 938.0mL. Mix thoroughly. Distribute into anaerobe tubes or bot-
tles. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and
anaerobically add per liter, 10.0mL sucrose solution, 10.0mL yeast ex-
tract solution, 10.0mL potassium nitrate solution, 10.0mL tryptone so-
lution, 10.0mL magnesium chloride solution, 10.0mL calcium chloride
solution, 1.0mL vitamin solution, and 1.0mL trace elements solution.
Mix thoroughly. Adjust pH to 7.0–7.5
Use:  For the cultivation of Oceanithermus profundus.