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Lactose Egg Yolk Milk Agar

[所属分类:培养基配方] [发布时间:2021-11-25] [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 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.

Measure 50.0mL of egg yolk emulsion and add to

50.0mL of 0.9% NaCl solution. Mix thoroughly. Fil

ter sterilize. Warm to 45°–50°C.

Inhibitor Solution:

Composition per 20.0mL:

Neomycin sulfate..............................................0.18g

NaN3 .................................................................0.24g

Preparation of Inhibitor Solution: Add neo

mycin sulfate and NaN3 to distilled/deionized water

and bring volume to 20.0mL. Mix thoroughly. Filter

sterilize.

Caution: Sodium azide is toxic. Azides also react

with metals and disposal must be highly diluted.

Preparation of Medium: Combine Columbia blood

agar base, lactose, agar, and Neutral Red and bring vol

ume to 1.0L. Adjust pH to 7.0. Autoclave for 15 min at

15 psi pressure–121°C. Cool to 55°C. Filter sterilize

skim milk. To 1.0L of cooled, sterile agar mixture, asep

tically 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 Pet

ri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Clostri

dium species.

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Lactose Egg Yolk Milk Agar