编号：
批号：

培养基配方

您现在的位置：网站首页 >> 培养基配方

Thermophilic Maintenance Medium

[所属分类:培养基配方] [发布时间:2021-9-7] [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:

NaHCO3 ........................................................................................ 3.0g

Yeast extract.................................................................................. 1.0g

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

KH2PO4......................................................................................... 0.4g

K2HPO4......................................................................................... 0.4g

MgSO4·7H2O ................................................................................ 0.1g

Cysteine-sulfide reducing solution ..........................................40.0mL

Fructose solution......................................................................25.0mL

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

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

Resazurin (0.01% solution)........................................................1.0mL

pH 5.6 ± 0.2 at 25°C

Cysteine-Sulfide Reducing Solution:

Composition per 100.0mL:

L-Cysteine·HCl·H2O ................................................................... 1.25g

Na2S·9H2O.................................................................................. 1.25g

Preparation of Cysteine-Sulfide Reducing Solution: Add L

cysteine·HCl·H2O and Na2S·9H2O to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly.

Fructose Solution:

Composition per 100.0mL:

Fructose....................................................................................... 20.0g

Preparation of Fructose Solution: Add fructose to distilled/de

ionized water and bring volume to 100.0mL. Mix thoroughly. Filter

sterilize.

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ........................................................................... 0.01g

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

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

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

Calcium pantothenate ................................................................5.0mg

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

Thioctic acid ..............................................................................5.0mg

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

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

Cyanocobalamin .....................................................................100.0μg

Preparation of Wolfe’s Vitamin Solution: Add components to

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

Wolfe’s Mineral Solution:

Composition per liter:

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

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

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

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

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

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

CaCl2............................................................................................. 0.1g

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

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

AlK(SO4)2·12H2O....................................................................... 0.01g

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

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

Preparation of Wolfe’s Mineral 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.

Preparation of Medium: Add components, except cysteine-sulfide

reducing solution and fructose solution, to distilled/deionized water

and bring volume to 935.0mL. Mix thoroughly. Gently heat and bring

to boiling. Continue boiling until resazurin turns colorless, indicating

reduction. Add 40.0mL of the cysteine-sulfide reducing solution. Au

toclave for 15 min at 15 psi pressure–121°C. Cool to 50°C under O2-

free 90% N2 + 10% CO2. Add 25.0mL of the sterile fructose solution.

Adjust the pH to 5.6 if necessary. Aseptically and anaerobically distrib

ute into sterile tubes. Cap with rubber stoppers.

Use: For the cultivation and maintenance of a variety of thermophilic

anaerobes, including Clostridium thermoautotrophicum.

在线客服

Thermophilic Maintenance Medium