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Urea Broth 10B for Ureaplasma urealyticum

[所属分类:培养基配方] [发布时间:2021-9-30] [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 100.5mL:

PPLO broth without Crystal Violet..........................................70.0mL

Horse serum, unheated.............................................................20.0mL

Fresh yeast extract solution .....................................................10.0mL

L-Cysteine·HCl·H2O solution.....................................................0.5mL

CVA enrichment ........................................................................0.5mL

Urea solution..............................................................................0.4mL

Phenol Red.................................................................................0.1mL

PPLO Broth without Crystal Violet:

Composition per 900.0mL:

Beef heart, solids from infusion.................................................. 16.1g

Peptone ....................................................................................... 3.25g

NaCl............................................................................................ 1.61g

Preparation of PPLO Broth without Crystal Violet: Add

components to distilled/deionized water and bring volume to 900.0mL.

Adjust pH to 5.5 with 2N HCl. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to 37°C.

Fresh Yeast Extract Solution:

Composition per 100.0mL:

Baker’s yeast live, pressed, starch-free....................................... 25.0g

Preparation of Fresh Yeast Extract Solution: Add the live Bak

er’s yeast to 100.0mL of distilled/deionized water. Autoclave for 90

min at 15 psi pressure–121°C. Allow to stand. Remove supernatant so

lution. Adjust pH to 6.6–6.8.

L-Cysteine·HCl·H2O Solution:

Composition per 50.0mL:

L-Cysteine·HCl·H2O ..................................................................... 1.0g

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cyste

ine·HCl·H2O to distilled/deionized water and bring volume to 50.0mL.

Mix thoroughly. Filter sterilize.

CVA Enrichment:

Composition per liter:

Glucose ..................................................................................... 100.0g

L-Cysteine·HCl·H2O.................................................................... 25.9g

L-Glutamine................................................................................. 10.0g

Adenine......................................................................................... 1.0g

L-Cystine·2HCl.............................................................................. 1.0g

Nicotinamide adenine dinucleotide ............................................ 0.25g

Cocarboxylase............................................................................... 0.1g

Guanine·HCl ............................................................................... 0.03g

Fe(NO3)3 ..................................................................................... 0.02g

Vitamin B12 ................................................................................. 0.01g

p-Aminobenzoic acid................................................................ 0.013g

Thiamine·HCl ............................................................................3.0mg

Preparation of CVA Enrichment: Add components to distilled/

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

ilize.

Urea Solution:

Composition per 30.0mL:

Urea............................................................................................... 3.0g

Preparation of Urea Solution: Add urea to distilled/deionized wa

ter and bring volume to 30.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Aseptically combine the components, ex

cept the PPLO broth without Crystal Violet. Aseptically add this mix

ture to the cooled, sterile PPLO broth without Crystal Violet. Mix

thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Ureaplasma urealyticum

and other Ureaplasma species. Urease-positive bacteria turn the

medium peach orange.

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Urea Broth 10B for Ureaplasma urealyticum