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培养基配方

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Thermotoga subterranea Medium

[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]

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............................................................................................ 12.0g

MgSO4·7H2O................................................................................ 0.5g

PIPES............................................................................................ 3.4g

KCl................................................................................................ 2.0g

Peptone ......................................................................................... 1.0g

Na2S·9H2O.................................................................................... 0.5g

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

NH4Cl ........................................................................................... 0.1g

CaCl2·2H2O .............................................................................. 0.025g

K2HPO4....................................................................................... 0.02g

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

Wolfe’s mineral solution..........................................................10.0mL

Wolfe’s vitamin solution..........................................................10.0mL

pH 7.0 ± 0.2 at 25°C

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ........................................................................ 10.0mg

p-Aminobenzoic acid................................................................. 5.0mg

Lipoic acid ................................................................................. 5.0mg

Nicotinic acid............................................................................. 5.0mg

Riboflavin .................................................................................. 5.0mg

Thiamine·HCl ............................................................................ 5.0mg

Calcium DL-pantothenate........................................................... 5.0mg

Biotin ......................................................................................... 2.0mg

Folic acid ................................................................................... 2.0mg

Vitamin B12 ................................................................................ 0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to

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

Filter sterilize.

Wolfe’s Mineral Solution:

Composition per liter:

MgSO4·7H2O................................................................................ 3.0g

Nitrilotriacetic acid ....................................................................... 1.5g

NaCl.............................................................................................. 1.0g

MnSO4·2H2O................................................................................ 0.5g

CoCl2·6H2O .................................................................................. 0.1g

ZnSO4·7H2O ................................................................................. 0.1g

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

FeSO4·7H2O.................................................................................. 0.1g

NiCl2·6H2O............................................................................... 0.025g

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

CuSO4·5H2O............................................................................... 0.01g

H3BO3 ......................................................................................... 0.01g

Na2MoO4·2H2O .......................................................................... 0.01g

Na2SeO3·5H2O........................................................................... 0.3mg

Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with

KOH. Add remaining components. Add distilled/deionized water to

1.0L. Adjust pH to 6.8.

Preparation of Medium: Prepare and dispense medium under

100% N2. Add components, except Wolfe’s vitamin solution, to dis

tilled/deionized water and bring volume to 990.0mL. Mix thoroughly.

Adjust pH to 7.0. Sparge with 100% N2. Autoclave for 15 min at 15 psi

pressure–121°C. Aseptically and anaerobically add 10.0mL of sterile

Wolfe’s vitamin solution. Mix thoroughly. Adjust medium pH to 7.5 by

adding sterile anaerobic 1N NaOH. Aseptically and anaerobically dis

tribute into sterile tubes or bottles.

Use: For the cultivation of Thermotoga subterranea.

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Thermotoga subterranea Medium