Lactose Egg Yolk Milk Agar

[所属分类:培养基配方] [发布时间:2021-4-28] [发布人:李佳颐] [阅读次数:] [返回]

Lactose Egg Yolk Milk Agar

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

Composition per 1206.0mL:

Lactose........................................................................................12.0g
Agar..............................................................................................1.0g
Columbia blood agar base .....................................................800.0mL
Skim milk...............................................................................150.0mL
Egg yolk emulsion, 50%..........................................................36.0mL
Inhibitor solution .....................................................................20.0mL
Neutral Red (1% solution).......................................................3.25mL

pH 7.0 ± 0.2 at 25°C

Columbia Blood Agar Base:
Composition per 800.0mL:
Agar ............................................................................................15.0g
Pantone........................................................................................10.0g
Bitone..........................................................................................10.0g
NaCl..............................................................................................5.0g
Tryptic digest of beef heart...........................................................3.0g
Cornstarch.....................................................................................1.0g
Preparation of Columbia Blood Agar Base: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Gently heat until boiling.
Egg Yolk Emulsion, 50%:
Composition per 100.0mL:
Chicken egg yolks............................................................................11
Whole chicken egg.............................................................................1
NaCl (0.9% solution)...............................................................50.0mL
Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100
dilution of saturated mercuric chloride solution for 1 min. Crack eggs
and separate yolks from whites. Mix egg yolks with 1 chicken egg.
Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add
to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize.
Warm to 45°–50°C.
Inhibitor Solution:
Composition per 20.0mL:
Neomycin sulfate........................................................................0.18g
NaN 3 ...........................................................................................0.24g
Caution: Sodium azide is toxic. Azides also react with metals and
disposal must be highly diluted.
Preparation of Inhibitor Solution: Add neomycin sulfate and
NaN 3 to distilled/deionized water and bring volume to 20.0mL. Mix
thoroughly. Filter sterilize.
Preparation of Medium: Combine Columbia blood agar base, lac-
tose, agar, and Neutral Red and bring volume to 1.0L. Adjust pH to 7.0.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 55°C. Filter ster-
ilize skim milk. To 1.0L of cooled, sterile agar mixture, aseptically add
150.0mL of sterile skim milk, 36.0mL of sterile egg yolk emulsion, 50%,
and 20.0mL of sterile inhibitor solution. Mix thoroughly. Pour into sterile
Petri dishes or distribute into sterile tubes.
Use: For the cultivation and maintenance of Clostridium species.