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U4 Medium

[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]

U4 Medium

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

Composition per 100.0mL:

Hartley’s digest broth...............................................................20.0mL

Fetal calf serum........................................................................15.0mL

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

Hanks’ balanced salt solution, 10X ...........................................4.0mL

MgSO4·5H2O (0.025% solution) ...............................................1.0mL

Urea (20% solution).................................................................0.25mL

Phenol Red (1% solution) ..........................................................0.2mL

pH 6.0–6.2 at 25°C

Hartley’s Digest Broth:

Composition per 10.0L:

Ox heart................................................................................... 3000.0g

Pancreatin.................................................................................... 50.0g

Na2CO3, anhydrous (0.8% solution).............................................5.0L

HCl, concentrated ....................................................................80.0mL

pH 7.5 ± 0.2 at 25°C

Preparation of Hartley’s Digest Broth: Finely mince the ox

heart. Add the meat to 5.0L of distilled/deionized water. Gently heat

and bring to 80°C. Add the 5.0L of Na2CO3 solution. Cool to 45°C.

Add pancreatin and maintain at 45°C for 4 hr while stirring. Add the

HCl and steam at 100°C for 30 min. Cool to room temperature. Adjust

pH to 8.0 with 1N NaOH. Gently heat and bring to boiling. Continue

boiling for 25 min. Filter while hot. Cool to room temperature. Adjust

pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°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.

Hanks’ Balanced Salt Solution, 10X:

Composition per liter:

Na2Cl........................................................................................... 80.0g

Glucose ....................................................................................... 10.0g

KCl................................................................................................ 4.0g

CaCl2............................................................................................. 1.4g

MgCl2·6H2O.................................................................................. 1.0g

MgSO4·7H2O ................................................................................ 1.0g

Na2HPO4·7H2O............................................................................. 0.9g

KH2PO4......................................................................................... 0.6g

Preparation of Hanks’ Balanced Salt Solution, 10X: Add

components to distilled/deionized water and bring volume to 100.0mL.

Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.0–6.2

with HCl. Filter-sterilize medium. Aseptically distribute into sterile

tubes or flasks.

Use: For the cultivation and maintenance of Ureaplasma diversum.

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U4 Medium