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Thermotoga petrophila Medium (DSMZ Medium 913)

[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]

Thermotoga petrophila Medium

(DSMZ Medium 913)

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

Composition per liter:

MOPS ........................................................................................... 5.0g

Yeast extract.................................................................................. 2.0g

(NH4)2SO4 .................................................................................... 1.0g

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

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

Artificial seawater..................................................................750.0mL

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

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

Na2S·9H2O solution.................................................................10.0mL

pH 7.0 ± 0.2 at 25°C

Artificial Seawater:

Composition per liter:

NaCl ............................................................................................ 27.7g

MgSO4·7H2O ................................................................................ 7.0g

MgCl2·6H2O.................................................................................. 5.5g

KCl.............................................................................................. 0.65g

NaBr.............................................................................................. 0.1g

H3BO3 ......................................................................................30.0mg

SrCl2·6H2O...............................................................................15.0mg

Citric acid.................................................................................10.0mg

KI ............................................................................................ 0.05mg

CaCl2·2H2O................................................................................ 2.25 g

Preparation of Artificial Seawater: Add components to distilled/

deionized water and bring volume to 1.0L. Mix thoroughly.

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.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.3g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

tilled/deionized water and bring volume to 10.0mL. Sparge with N2.

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

aerobically.

Preparation of Medium: Add components, except vitamin solution

and Na2S·9H2O solution, to 750.0mL artificial seawater. Bring volume

to 980.0mL with distilled/deionized water. Mix thoroughly. Gently heat

and bring to boiling. Boil for 3 min. Cool to room temperature while

sparging with 100% N2. Distribute under 100% N2 into anaerobe tubes

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

and anaerobically add, per liter of medium, 10.0mL vitamin solution

and 10.0mL Na2S·9H2O solution. Mix thoroughly. The final pH of the

medium should be 7.0.

Use: For the cultivation of Thermotoga petrophila and Thermotoga

naphthophila.

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Thermotoga petrophila Medium (DSMZ Medium 913)