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培养基配方

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Spirochaeta caldaria Medium

[所属分类:培养基配方] [发布时间:2021-8-23] [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:

Glucose ......................................................................................... 2.0g

Pancreatic digest of casein............................................................ 2.0g

L-Cysteine ................................................................................... 0.25g

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

Wolfe’s mineral solution ..........................................................25.0mL

Trace elements solution SL-10 ................................................10.0mL

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

pH 7.0 ± 0.2 at 25°C

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. Filter sterilize. Sparge with 100% N2.

Trace Elements Solution SL-10:

Composition per liter:

FeCl2·4H2O................................................................................... 1.5g

CoCl2·6H2O ........................................................................... 190.0mg

MnCl2·4H2O .......................................................................... 100.0mg

ZnCl2........................................................................................ 70.0mg

Na2MoO4·2H2O ....................................................................... 36.0mg

NiCl2·6H2O.............................................................................. 24.0mg

H3BO3 ........................................................................................ 6.0mg

CuCl2·2H2O ............................................................................... 2.0mg

HCl (25% solution)..................................................................10.0mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O

to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized

water and bring volume to 1.0L. Add remaining components. Mix thor

oughly. Filter sterilize. Sparge with 100% N2.

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. Sparge with 100% N2.

Preparation of Medium: Prepare and dispense medium under

100% N2. Add components, except Wolfe’s mineral solution, trace el

ements SL-10 solution, and Wolfe’s vitamin solution, to distilled/de

ionized water and bring volume to 955.0mL. Mix thoroughly. Sparge

with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Asep

tically and anaerobically add 25.0mL of sterile Wolfe’s mineral solu

tion, 10.0mL of sterile trace elements SL-10 solution, and 10.0mL of

sterile Wolfe’s vitamin solution. Mix thoroughly. Aseptically and an

aerobically distribute into sterile tubes or flasks.Adjust pH to 7.0 with

sterile NaOH.

Use: For the cultivation of Spirochaeta caldaria

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Spirochaeta caldaria Medium