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U9 Broth with Amphotericin B

[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 101.6mL:

U9 base ....................................................................................95.0mL

Horse serum, unheated...............................................................5.0mL

Antibiotic solution .....................................................................1.0mL

Urea solution..............................................................................0.5mL

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

pH 6.0 ± 0.2 at 25°C

U9 Base:

Composition per 100.0mL:

NaCl............................................................................................ 0.63g

Pancreatic digest of casein........................................................ 0.425g

Papaic digest of soybean meal.................................................. 0.075g

K2HPO4..................................................................................... 0.063g

Glucose ..................................................................................... 0.063g

KH2PO4....................................................................................... 0.02g

Preparation of U9 Base: Add components to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Adjust pH to 5.5

with 1N HCl. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

45°–50°C.

Antibiotic Solution:

Composition per 10.0mL:

Penicillin G ................................................................................. 0.63g

Amphotericin B ......................................................................... 2.5mg

Preparation of Antibiotic Solution: Add penicillin G and ampho

tericin B to distilled/deionized water and bring volume to 10.0mL. Mix

thoroughly. Filter sterilize.

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.

Phenol Red Solution:

Composition per 10.0mL:

Phenol Red.................................................................................... 0.1g

Preparation of Phenol Red Solution: Add Phenol Red to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Filter sterilize.

Preparation of Medium: To 95.0mL of cooled, sterile U9 base,

aseptically add 5.0mL of sterile horse serum, 1.0mL of sterile antibiot

ic solution, 0.5mL of sterile urea solution, and 0.1mL of sterile Phenol

Red solution. Mix thoroughly. Aseptically distribute into sterile tubes

or flasks.

Use: For the isolation and identification of T-strain mycoplasmas from

clinical specimens, especially Ureaplasma urealyticum. T-mycoplas

mas are the only members of the Mycoplasma group known to contain

urease. Bacteria with urease activity turn the medium dark pink.

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U9 Broth with Amphotericin B