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Thermincola Medium (DSMZ Medium 1028)

[所属分类:培养基配方] [发布时间:2021-9-6] [发布人:网站管理员2] [阅读次数:] [返回]

Thermincola Medium

(DSMZ Medium 1028)

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

Composition per liter:

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

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

KH2PO4......................................................................................... 0.5g

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

Na-acetate ..................................................................................... 0.2g

CaCl2·2H2O .................................................................................. 0.1g

Resazurin .................................................................................. 0.5mg

Vitamin solution.......................................................................20.0mL

Bicarbonate solution ................................................................10.0mL

Carbonate solution...................................................................10.0mL

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

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

Wolfe's mineral elixir.................................................................1.0mL

pH 8.0 ± 0.2 at 25°C

Sulfide Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 1.0g

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.

Bicarbonate Solution:

Composition per 10.0mL:

NaHCO3 ........................................................................................ 0.5g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge

with gas mixture of 80% N2 + 20% CO2. Filter sterilize.

Carbonate Solution:

Composition per 10.0mL:

Na2CO3 ......................................................................................... 0.5g

Preparation of Carbonate Solution: Add Na2CO3 to distilled/de

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

with gas mixture of 80% N2 + 20% CO2. Filter sterilize.

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.

Sparge with 100% N2 gas. Autoclave for 15 min at 15 psi pressure–

121°C.

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 components one

at a time. Mix thoroughly to dissolve.

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.

Preparation of Medium: Add components, except vitamin solu

tion, to distilled/deionized water and bring volume to 950.0mL. Mix

thoroughly. Gently heat and bring to boiling. Boil for 1 min. Cool to

room temperature while sparging with 100% N2 gas. Dispense into cul

ture vessels under an atmostphere of 100% CO (carbon monoxide) .

Culture vessels should be filled to approximately 20%. Autoclave for

15 min at 15 psi pressure–121°C. Cool to room temperature. Aspetical

ly add carbonate, bicarbonate, vitamin, and sulfide solutions. Mix thor

oughly. Adjust the pH to 8.0 with sterile, anoxic 1N HCl. Prior to

inoculation aseptically add yeast extract solution.

Use: For the cultivation of Thermincola carboxydiphila.

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Thermincola Medium (DSMZ Medium 1028)