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Base Layer Agar with Nutrient Overlay Agar

[所屬分類:培養基配方] [發布時間:2021-11-3] [發布人:網站管理員2] [閱讀次數:] [返回]
Base Layer Agar with
Nutrient Overlay Agar

山東拓普生物工程有限公司 培養基配方 //ljxinglong.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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