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U Broth (Ureaplasma Broth)

[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]

U Broth

(Ureaplasma Broth)

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

Composition per 99.5mL:

Base agar..................................................................................65.0mL

Horse serum.............................................................................20.0mL

Yeast dialysate .........................................................................10.0mL

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

MES (2-N-morpholinoethane

sulfonic acid) buffer solution ..............................................1.0mL

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

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

pH 5.5 ± 0.2 at 25°C

Base Agar:

Composition per liter:

Papaic digest of soybean meal.................................................... 20.0g

Agarose....................................................................................... 10.0g

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

Phenol Red (2% solution)..........................................................1.0mL

Preparation of Base Agar: Add components to distilled/deionized

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

to boiling. Adjust pH to 7.3. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to 45°–50°C.

Yeast Dialysate:

Composition per 10.0mL:

Yeast, active dried..................................................................... 450.0g

Preparation of Yeast Dialysate: Add active, dried yeast to dis

tilled/deionized water and bring volume to 1250.0mL. Gently heat and

bring to 40°C. Autoclave for 15 min at 15 psi pressure–121°C. Put into

dialysis tubing. Dialyze against 1.0L of distilled/deionized water for 2

days at 4°C. Discard tubing and its contents. Autoclave dialysate for 15

min at 15 psi pressure–121°C. Store at −20°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.

Preparation of Medium: To 65.0mL of cooled, sterile base agar,

aseptically add 10.0mL of sterile yeast dialysate, 20.0mL of horse se

rum, 2.0mL of sterile penicillin solution, 1.0mL of sterile MES buffer

solution, 1.0mL of sterile Na2SO3 solution, and 0.5mL of sterile urea

solution. Mix thoroughly. Pour into 10mm × 35mm Petri dishes in

5.0mL volumes. Allow plates to stand overnight at 25°C to remove ex

cess surface moisture. Use within 48 hr.

Use: For the isolation and cultivation of Ureaplasma urealyticum.

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U Broth (Ureaplasma Broth)