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Base Layer Agar with Nutrient Overlay Agar

[所属分类:培养基配方] [发布时间:2021-11-3] [发布人:网站管理员2] [阅读次数:] [返回]

Base Layer Agar with

Nutrient 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 2.5L:

Fat substrate......................................................50.0g

Nutrient agar ...................................................... 1.5L

Basal medium .................................................... 1.0L

Fat Substrate:

Composition:

Fat .....................................................................50.0g

Preparation of Fat Substrate: Tributyrin, corn

oil, soybean oil, any cooking oil, lard, tallow, or trig

lycerides that do not contain antioxidants or other in

hibitory substances may be used. Remove free fatty

acids in the fat substrate by dissolving 50.0g of fat

substrate in 500.0mL of petroleum ether. Pass the so

lution through an activated alumina column. Remove

the petroleum ether by evaporation on a steam table

under 100% N2. Autoclave for 30 min at 15 psi pres

sure–121°C. Cool to 50°C.

Nutrient Agar:

Composition per liter:

Agar ..................................................................15.0g

Pancreatic digest of gelatin.................................5.0g

Beef extract.........................................................3.0g

Preparation of Nutrient Agar: Add compo

nents to distilled/deionized water and bring volume

to 1.0L. Mix thoroughly. Gently heat while stirring

and bring to boiling. Distribute into tubes or flasks.

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

to 45°–50°C.

Source: The medium is available as a premixed

powder from BD Diagnostic Systems.

Basal Medium:

Composition per liter:

Agar ..................................................................15.0g

Victoria Blue B solution ............................. 200.0mL

Preparation of Basal Medium: Add agar to

800.0mL of distilled/deionized water. If tributyrin is

used as the fat substrate, add agar to 1.0L of distilled/

deionized water. Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to 50°C. If tributyrin is not used as

the fat substrate, aseptically add 200.0mL of Victoria

Blue B solution. Mix thoroughly.

Victoria Blue B Solution:

Composition per 200.0mL:

Victoria Blue B .................................................0.12g

Preparation of Victoria Blue B Solution: Add

the Victoria Blue B solution to 200.0mL of distilled/

deionized water. Mix thoroughly. Filter sterilize.

Warm to 50°C.

Preparation of Medium: Aseptically combine

1.0L of sterile basal medium with 50.0g of sterile fat

substrate in a warm, sterile blender container. Blend

for 1 min until homogenized. Rapidly pour into ster

ile Petri dishes in 7.0mL volumes. Dry the surface of

the plates by partially opening the lids in a laminar

flow hood for 15 min. Add dilution of food samples

to be tested. When the inoculum is dry, pour nutrient

agar as an overlay onto each plate. Use 10–12mL of

nutrient agar per plate.

Use: For the isolation, cultivation, and identification

of lipolytic microorganisms from food.

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Base Layer Agar with Nutrient Overlay Agar