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Vulcanibacillus Medium (DSMZ Medium 1042)

[所屬分類:培養基配方] [發布時間:2021-10-12] [發布人:網站管理員2] [閱讀次數:] [返回]
Vulcanibacillus Medium
(DSMZ Medium 1042)

山東拓普生物工程有限公司 培養基配方 //ljxinglong.com

Composition per liter:

Sea salts, Sigma .......................................................................... 30.0g
NaNO3........................................................................................... 1.0g
Na-pyruvate ................................................................................. 1.0g
Resazurin ..................................................................................0.5mg
VoSO4·2H2O ............................................................................0.05mg
Vitamin solution.......................................................................20.0mL
Yeast extract solution ...............................................................10.0mL
Bicarbonate solution ................................................................10.0mL
Iron sulfate solution .................................................................10.0mL
Wolfe's mineral elixir.................................................................2.0mL
pH 6.9 ± 0.2 at 25°C
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract ................................................................................. 2.0g
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 room temper
ature. Sparge with 100% N2.
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 B12 ................................................................................ 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.
Wolfe’s Mineral Elixir:
Composition per liter:
MgSO4·7H2O.............................................................................. 30.0g
NaCl............................................................................................ 10.0g
MnSO4·2H2O................................................................................ 5.0g
(NH4)2NiSO4·6H2O ...................................................................... 2.8g
CoCl2·6H2O .................................................................................. 1.8g
ZnSO4·7H2O ................................................................................. 1.8g
FeSO4·7H2O.................................................................................. 1.0g
CaCl2·2H2O .................................................................................. 1.0g
KAl(SO4)2·12H2O....................................................................... 0.18g
CuSO4·5H2O................................................................................. 0.1g
H3BO3 ........................................................................................... 0.1g
Na2MoO4·2H2O ............................................................................ 0.1g
Na2SeO4 ........................................................................................ 0.1g
Na2WO4·2H2O.............................................................................. 0.1g
Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of dis
tilled/deionized water to 1.0 with dilute H2SO4. Add remaining com
ponents one at a time. Mix thoroughly to dissolve.
Bicarbonate Solution:
Composition per 10.0mL:
NaHCO3........................................................................................ 1.0g
Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with a gas mixture of 80% N2 + 20% CO2. Filter sterilize.
Iron Sulfate Solution:
Composition per 10.0mL:
FeSO4·7H2O.................................................................................. 0.1g
Preparation of Iron Sulfate Solution: Add FeSO4·7H2O to 0.1N
sulfuric acid and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2. Filter sterilize.
Preparation of Medium: Add components, except iron sulfate, bi
carbonate, yeast extract, and vitamin solutions, to distilled/deionized
water and bring volume to 940.0mL. Mix thoroughly. Adjust pH to 7.0.
Gently heat and bring to boiling. Boil for 1 min. Cool to room temper
ature while sparging with 100% N2. Dispense into culture vessels un
der an atmosphere of 100% N2. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to room temperature. Aseptically add iron sul
fate, bicarbonate, yeast extract, and vitamin solutions. Mix thoroughly.
Adjust pH to 6.8–7.0. Aseptically dispense into culture vessels. It may
be necessary to add 10–20mg sodium dithionite per liter (e.g., from a
5% (w/v) solution, freshly prepared under N2 and filter-sterilized), if
the medium is not completely reduced after inoculation.
Use: For the cultivation of Clostridiisalibacter paucivoran.
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