培养基配方

您现在的位置：网站首页 >> 培养基配方

Steenken and Smith Agar (Hohn’s Medium, Modified)

[所属分类:培养基配方] [发布时间:2021-8-26] [发布人:网站管理员2] [阅读次数:] [返回]

Steenken and Smith Agar

(Hohn’s Medium, Modified)

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

Composition per 2065.0mL:

Homogenized whole egg .....................................................1500.0mL

Stock salts solution ................................................................500.0mL

Lacmoid solution .....................................................................25.0mL

HCl (1N solution) ....................................................................40.0mL

pH 6.6 ± 0.2 at 25°C

Stock Salts Solution:

Composition per 500.0mL:

KH2PO4......................................................................................... 2.0g

Asparagine .................................................................................... 1.5g

Magnesium citrate ..................................................................... 1.25 g

Na2HPO4, anhydrous ................................................................... 1.2 g

MgSO4 .......................................................................................... 0.3g

Glycerol ...................................................................................60.0mL

Preparation of Stock Salts Solution: Add components, except

glycerol, to distilled/deionized water that has been warmed to 80°C.

Bring volume to 440.0mL. Mix thoroughly. Add 60.0mL of glycerol.

Mix thoroughly. Autoclave for 20 min at 10 psi pressure–115°C. Cool

to 25°C. Aseptically divide the solution into two 250.0mL parts.

Lacmoid Solution:

Composition per 100.0mL:

Lacmoid ........................................................................................ 1.0g

Ethanol (50% solution)..........................................................100.0mL

Preparation of Lacmoid Solution: Add lacmoid to 100.0mL of

ethanol solution. Mix thoroughly.

Preparation of Medium: To one 250.0mL part of sterile stock salts

solution, add 25.0mL of lacmoid solution. Mix thoroughly. To the other

250.0mL part of sterile stock salts solution, add 40.0mL of HCl solution.

Mix thoroughly. Soak eggs in 70% ethanol for 10 min. Dry between ster

ile towels. Break eggs into a sterile container. Aseptically homogenize

the whole eggs with a sterile glass rod. Add both stock salt solutions to

the homogenized whole egg mixture. Mix thoroughly. Filter through

sterile cheesecloth. Aseptically distribute into sterile tubes. Inspissate

medium at 85°C (moist heat) for 90 min on 2 consecutive days.

Use: For the cultivation and maintenance of Mycobacterium microti.