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Thermoanaerobium brockii Medium

[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]

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

Composition per liter:

Pancreatic digest of casein.......................................................... 10.0g

Yeast extract.................................................................................. 3.0g

K2HPO4......................................................................................... 1.5g

NH4Cl ........................................................................................... 0.9g

NaCl.............................................................................................. 0.9g

KH2PO4....................................................................................... 0.75g

MgCl2·6H2O ................................................................................. 0.2g

Glucose solution ......................................................................25.0mL

Na2S·9H2O (10% solution)......................................................10.0mL

Trace elements solution .............................................................9.0mL

Wolfe’s Vitamin solution ...........................................................5.0mL

Resazurin (0.025% solution)......................................................4.0mL

FeSO4·7H2O (10% solution)....................................................0.03mL

pH 7.3 ± 0.2 at 25°C

Glucose Solution:

Composition per 100.0mL:

Glucose ....................................................................................... 20.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster

ilize.

Trace Elements Solution:

Composition per liter:

Nitrilotriacetic acid ..................................................................... 12.5g

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

FeCl3·4H2O ................................................................................... 0.2g

MnCl2·4H2O.................................................................................. 0.1g

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

ZnCl2............................................................................................. 0.1g

CuCl2........................................................................................... 0.02g

Na2SeO3 ...................................................................................... 0.02g

CoCl2·6H2O .............................................................................. 0.017g

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

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

Preparation of Trace Elements 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.

Wolfe’s Vitamin Solution:

Composition per liter:

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

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.

Preparation of Medium: Add components, except glucose solu

tion, to distilled/deionized water and bring volume to 975.0mL. Mix

thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. While still

hot, aseptically add 25.0mL of the sterile glucose solution under 97%

N2 + 3% H2. Adjust pH to 7.3 if necessary. Aseptically and anaerobi

cally distribute into tubes. Cap with rubber stoppers.

Use: For the cultivation and maintenance of Thermoanaerobium

brockii.

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Thermoanaerobium brockii Medium