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Thermovenabulum Medium (DSMZ Medium 962)

[所属分类:培养基配方] [发布时间:2021-9-8] [发布人:网站管理员2] [阅读次数:] [返回]

Thermovenabulum Medium

(DSMZ Medium 962)

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

Composition per liter:

NaHCO3 ........................................................................................ 0.7g

NH4Cl ......................................................................................... 0.33g

KH2PO4....................................................................................... 0.33g

MgCl2·6H2O................................................................................ 0.33g

CaCl2·2H2O................................................................................. 0.33g

KCl.............................................................................................. 0.33g

Yeast extract................................................................................ 0.05g

Meat extract solution ...............................................................50.0mL

Sodium fumarate solution........................................................50.0mL

Vitamin solution.......................................................................10.0mL

Trace elements solution ...........................................................10.0mL

Selenite-tungstate solution.........................................................1.0mL

pH 7.0 ± 0.2 at 25°C

Selenite-Tungstate Solution

Composition per liter:

NaOH ............................................................................................ 0.5g

Na2WO4·2H2O ...........................................................................4.0mg

Na2SeO3·5H2O...........................................................................3.0mg

Preparation of Selenite-Tungstate Solution: Add components

to distilled/deionized water and bring volume to 1.0L. 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 B12 ................................................................................ 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.

Sodium Fumarate Solution:

Composition per 50.0mL:

Na2-fumarate............................................................................... 3.20g

Preparation of Sodium Fumarate Solution: Add Na2-fumarate

to distilled/deionized water and bring volume to 50.0mL. Sparge with

N2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Meat Extract Solution:

Composition per 50.0mL:

Meat extract .................................................................................. 3.0g

Preparation of Meat Extract Solution: Add meat extract to dis

tilled/deionized water and bring volume to 50.0mL. Sparge with N2.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Preparation of Medium: Add components, except NaHCO3, vita

min solution, sodium fumarate solution, and meat extract solution, to

distilled/deionized water and bring volume to 890.0mL. Mix thorough

ly. Gently heat and bring to boiling. Boil for 3 min. Cool to 25°C while

sparging with 80% N2 + 20% CO2. Add solid NaHCO3. Adjust pH to

6.8–7.0. Distribute to anaerobe tubes or bottles under 80% N2 + 20%

CO2. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and

anaerobically add, per liter of medium, 10.0mL sterile vitamin solu

tion, 50.0mL sterile meat extract solution, and 50.0mL sterile sodium

fumarate solution. Mix thoroughly. The final pH should be 6.5.

Use: For the cultivation of Thermovenabulum ferriorganovorum.

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Thermovenabulum Medium (DSMZ Medium 962)