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Urea Test Broth

[所属分类:培养基配方] [发布时间:2021-10-8] [发布人:网站管理员2] [阅读次数:] [返回]

Urea Test Broth

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

Composition per 99.6mL:

H broth base.............................................................................85.0mL

Horse serum.............................................................................10.0mL

Penicillin solution ......................................................................2.0mL

MES (2-N-morpholinoethane sulfonic acid) buffer solution.....1.0mL

Na2SO3 solution.........................................................................1.0mL

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

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

pH 7.2 ± 0.2 at 25°C

H Broth Base:

Composition per liter:

NaCl.............................................................................................. 5.0g

Pancreatic digest of casein............................................................ 5.0g

Peptone ......................................................................................... 5.0g

Beef extract................................................................................... 3.0g

K2HPO4......................................................................................... 2.5g

Glucose ......................................................................................... 1.0g

Preparation of H Broth Base: Add components to distilled/deion

ized water and bring volume to 1.0L. Mix thoroughly. Gently heat and

bring to boiling. Distribute into tubes in 4.0mL volumes. Autoclave for

15 min at 10 psi pressure–115°C. Cool to 45°–50°C.

Penicillin Solution:

Composition per 10.0mL:

Penicillin.............................................................................. 100,000U

Preparation of Penicillin Solution: Add penicillin to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster

ilize.

MES Buffer Solution:

Composition per 100.0mL:

MES (2-N-morpholinoethane sulfonic acid) buffer.................. 19.52g

Preparation of MES Buffer Solution: Add MES buffer to dis

tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.

Adjust pH to 5.5. Filter sterilize.

Na2SO3 Solution:

Composition per 10.0mL:

Na2SO3 ...................................................................................... 0.126g

Preparation of Na2SO3 Solution: Add Na2SO3 to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril

ize.

Urea Solution:

Composition per 100.0mL:

Urea............................................................................................... 6.0g

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

ter and bring volume to 100.0mL. 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 85.0mL of cooled sterile H broth base,

aseptically add 10.0mL of sterile horse serum, 2.0mL of sterile penicil

lin solution, 1.0mL of MES buffer solution, 1.0mL of Na2SO3 solution,

0.5mL of urea solution, and 0.1 mL of sterile Phenol Red solution. Mix

thoroughly. Aseptically distribute into test tubes in 3.0mL volumes.

Use: For the cultivation and differentiation of Ureaplasma species

based on their production of urease.

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Urea Test Broth