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Pork Plasma Fibrinogen Overlay Agar

[所属分类:培养基配方] [发布时间:2021-12-10] [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 plate:

Baird-Parker agar, modified.......................... 15.0mL

Pork plasma fibrinogen overlay agar .............. 8.0mL

pH 7.0 ± 0.1 at 25°C

Baird-Parker Agar, Modified:

Composition per liter:

Agar ..................................................................17.0g

Glycine..............................................................12.0g

Sodium pyruvate...............................................10.0g

Pancreatic digest of casein................................10.0g

Beef extract.........................................................5.0g

LiCl .....................................................................5.0g

Yeast extract........................................................1.0g

Chapman tellurite solution.............................. 1.0mL

pH 7.0 ± 0.2 at 25°C

Chapman Tellurite Solution:

Composition per 100.0mL:

K2TeO3 ................................................................1.0g

Preparation of Chapman Tellurite Solution:

Add K2TeO3 to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Baird-Parker Agar, Modi

fied: Add components, except Chapman tellurite solu

tion, to distilled/deionized water and bring volume to

999.0mL. Mix thoroughly. Gently heat and bring to

boiling. Autoclave for 15 min at 15 psi pressure–121°C.

Cool to 45–50°C. Aseptically add 1.0mL of sterile

Chapman tellurite solution. Mix thoroughly but gently.

Pork Plasma Fibrinogen Overlay Agar:

Composition per 100.5mL:

Agar solution................................................. 50.0mL

Bovine fibrinogen solution ........................... 47.5mL

Pork plasma..................................................... 2.5mL

Trypsin inhibitor solution ............................... 0.5mL

Agar Solution:

Composition per 50.0mL:

Agar ....................................................................0.7g

Preparation of Agar Solution: Add agar to dis

tilled/deionized water and bring volume to 50.0mL.

Mix thoroughly. Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to 45°–50°C.

Bovine Fibrinogen Solution:

Composition per 50.0mL:

Bovine fibrinogen, fraction I ..............................0.4g

Sodium phosphate buffer

(0.05M solution, pH 7.0) ........................ 50.0mL

Preparation of Bovine Fibrinogen Solution:

Grind bovine fibrinogen in a mortar to a fine powder.

Add bovine fibrinogen to 50.0mL of sodium phos

phate buffer. Mix thoroughly on a magnetic stirrer for

30 min. Filter through Whatman #1 filter paper. Filter

sterilize.

Pork Plasma:

Composition per 10.0mL:

Pork plasma-EDTA....................................... 10.0mL

Preparation of Pork Plasma: Filter sterilize

fresh or rehydrated commercial pork plasma-EDTA.

Trypsin Inhibitor Solution:

Composition per 5.0mL:

Trypsin inhibitor .............................................0.015g

Sodium phosphate buffer

(0.05M solution, pH 7.0) .......................... 5.0mL

Preparation of Trypsin Inhibitor Solution:

Add trypsin inhibitor to 5.0mL of sodium phosphate

buffer. Mix thoroughly. Filter sterilize.

Preparation of Pork Plasma Fibrinogen

Overlay Agar: Aseptically combine 50.0mL of

cooled, sterile agar solution, 47.5mL of sterile bovine

fibrinogen solution, 2.5mL of sterile pork plasma,

and 0.5mL of trypsin inhibitor solution. Mix thor

oughly. Maintain at 45°–50°C but use within 1 hr.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Pour cooled, sterile

Baird-Parker agar, modified, into sterile Petri dishes

in 15.0mL volumes. Allow agar to solidify. Overlay

each plate with 8.0mL of sterile pork plasma fibrino

gen overlay agar.

Use: For the cultivation of Staphylococcus aureus

from foods.

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Pork Plasma Fibrinogen Overlay Agar