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Spirochaeta americana Medium (DSMZ Medium 1165)

[所属分类:培养基配方] [发布时间:2021-8-20] [发布人:网站管理员2] [阅读次数:] [返回]

Spirochaeta americana Medium

(DSMZ Medium 1165)

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

NaHCO3 ..................................................................................... 24.0g

Na2CO3 ....................................................................................... 2.76g

NH4Cl .......................................................................................... 1.0g

KCl ............................................................................................... 0.2g

K2HPO4......................................................................................... 0.2g

MgCl2·6H2O.................................................................................. 0.1g

Resazurin ..................................................................................1.0mg

Sulfide solution ........................................................................10.0mL

Glucose solution ......................................................................10.0mL

Yeast extract solution...............................................................10.0mL

Vitamin solution.........................................................................2.0mL

Trace elements solution .............................................................1.0mL

pH 9.4 ± 0.2 at 25°C

Yeast Extract Solution:

Composition per 10.0mL:

Yeast extract ................................................................................. 0.5g

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.

Glucose Solution:

Composition per 10.0mL:

D-Glucose...................................................................................... 5.0g

Preparation of Glucose Solution: Add D-glucose to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster

ilize.

Sulfide Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.4g

Preparation of Sulfide Solution: Add Na2S·9H2O to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave

under 100% N2 for 15 min at 15 psi pressure–121°C. Cool to room

temperature.

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.

Trace Elements Solution:

Composition per 200.0mL:

MnCl2·4H2O ............................................................................... 0.72g

Fe(NH4)2(SO4)2·6H2O ................................................................. 0.4g

FeSO4·7H2O.................................................................................. 0.2g

CoCl2·6H2O .................................................................................. 0.2g

ZnSO4·7H2O ................................................................................. 0.2g

NiCl2·6H2O................................................................................... 0.1g

Na2MoO4·2H2O ......................................................................... 0.02g

CuSO4·5H2O............................................................................... 0.02g

H3BO3 ......................................................................................... 0.02g

KAl(SO4)2·12H2O....................................................................... 0.02g

HCl.............................................................................................5.0mL

Preparation of Trace Elements Solution: Add components to dis

tilled/deionized water and bring volume to 200.0mL. Mix thoroughly.

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

Preparation of Medium: Add components, except carbonate, bi

carbonate, sulfide solution, yeast extract solution, glucose solution,

and vitamin solution, to distilled/deionized water and bring volume to

968.0mL. Mix thoroughly. Gently heat and bring to boiling. Cool to

room temperature while sparging with 100% N2 gas. Add the Na2CO3

and NaHCO3. Mix thoroughly while sparging with 100% N2 gas. Ad

just pH to 9.4. Dispense into culture vessels (Hungate tubes or serum

bottles). Autoclave for 15 min at 15 psi pressure–121°C. Cool to room

temperature. Aseptically add sulfide solution, yeast extract solution,

glucose solution, and vitamin solution. Mix thoroughly.

Use: For the cultivation of Spirochaeta americana.

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Spirochaeta americana Medium (DSMZ Medium 1165)