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Stonebrink’s Medium

[所属分类:培养基配方] [发布时间:2021-12-17] [发布人:网站管理员2] [阅读次数:] [返回]

Stonebrink’s Medium

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

Composition per 1600.0mL:

Homogenized whole egg ................................... 2.0L

Mineral salts solution......................................... 1.0L

Malachite green solution............................... 40.0mL

Mineral Salts Solution:

Na-pyruvate ......................................................12.5g

KH2PO4...............................................................7.0g

Na2HPO4·7H2O...................................................4.0g

Preparation of Mineral Salts Solution: Add

components to distilled/deionized water and bring

volume to 1.0L. Mix thoroughly. Autoclave for 15

min at 15 psi pressure–121°C. Cool to 50°C.

Malachite Green Solution:

Composition per 100.0mL:

Malachite green ..................................................2.0g

Preparation of Malachite Green Solution:

Add malachite to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly. Autoclave for

15 min at 15 psi pressure–121°C. Cool to 50°C.

Homogenized Whole Egg:

Composition per 2.0L:

Whole eggs ...................................................... 36-48

Preparation of Homogenized Whole Egg:

Use fresh eggs, less than 1 week old. Scrub the shells

with soap. Let stand in a soap solution for 30 min.

Rinse in running water. Soak eggs in 70% ethanol for

15 min. Break the eggs into a sterile container. Ho

mogenize by shaking. Filter through four layers of

sterile cheesecloth into a sterile graduated cylinder.

Measure out 2.0L.

Preparation of Medium: Aseptically add

40.0mL sterile malachite to 1.0L of sterile mineral

salts solution. Mix thoroughly. Aseptically add 2.0L

of homogenized whole egg. Mix thoroughly. Distrib

ute into sterile screw-capped tubes. Place tubes in a

slanted position. Inspissate at 85°C (moist heat) for

45 min.

Use: For the cultivation of Mycobacterium species,

especially bovine strains. For the primary isolation of

Mycobacterium bovis.