Pork Plasma Fibrinogen Overlay Agar

[所属分类:培养基配方] [发布时间:2021-7-9] [发布人:网站管理员2] [阅读次数:] [返回]

Pork Plasma Fibrinogen Overlay Agar

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:
K 2 TeO 3 ..........................................................................................1.0g
Preparation of Chapman Tellurite Solution: Add K 2 TeO 3 to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Filter sterilize.
Preparation of Baird-Parker Agar, Modified: Add compo-
nents, except Chapman tellurite solution, to distilled/deionized water and
bring volume to 999.0mL. Mix thoroughly. Gently heat and bring to boil-
ing. 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 thor-
oughly 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 distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–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 fi-
brinogen in a mortar to a fine powder. Add bovine fibrinogen to
50.0mL of sodium phosphate buffer. Mix thoroughly on a magnetic
stirrer for 30 min. Filter through Whatman #1 filter paper. Filter steril-
ize.
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 inhibi-
tor to 5.0mL of sodium phosphate buffer. Mix thoroughly. Filter steril-
ize.
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 thoroughly. 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.