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培养基配方

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Coletsos Selective Medium

[所属分类:培养基配方] [发布时间:2021-11-12] [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 1625mL:

Potato starch......................................................10.0g

Gelatin.................................................................4.0g

Asparagine ........................................................2.25g

KH2PO4...............................................................1.5g

Na-glutamate.......................................................1.0g

Na-pyruvate ........................................................1.0g

Mg-citrate .......................................................0.375g

Litmus...............................................................0.25g

Malachite green ................................................0.25g

MgSO4 ..............................................................0.15g

Activated carbon.................................................0.1g

Oligonucleotide mixture ..................................3.0mg

Homogenized egg ...................................... 625.0mL

Glycerol .......................................................... 7.5mL

Nalidixic acid solution.................................... 1.0mL

Lincomycin solution ....................................... 1.0mL

Cycloheximide solution.................................. 1.0mL

Nalidixic Acid Solution:

Composition per 100.0mL:

Nalidixic acid......................................................0.5g

Preparation of Nalidixic Acid Solution: Add

nalidixic acid to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly. Filter sterilize.

Cycloheximide Solution:

Composition per 100.0mL:

Cycloheximide....................................................1.5g

Ethanol.......................................................... 40.0mL

Preparation of Cycloheximide Solution: Add

cycloheximide to 40.0mL of ethanol. Mix thorough

ly. Bring volume to 100.0mL with distilled/deionized

water. Filter sterilize.

Lincomycin Solution:

Composition per 100.0mL:

Lincomycin.........................................................0.5g

Preparation of Lincomycin Solution: Add lin

comycin to distilled/deionized water and bring vol

ume to 100.0mL. Mix thoroughly. Filter sterilize.

Egg Mixture Solution:

Composition per liter:

Whole eggs ......................................................18–24

Preparation of Egg Mixture Solution: Use

fresh eggs, less than 1 week old. Scrub the shells with

soap. Let stand in a soap solution for 30 min. Rinse

in running water. Soak eggs in 70% ethanol for 15

min. Break the eggs into a sterile container. Separate

egg whites from egg yolks. Combine 8 parts egg

white with 2 parts egg yolk. Homogenize by shaking.

Filter through four layers of sterile cheesecloth into a

sterile graduated cylinder. Measure out 1.0L.

Preparation of Medium: Add glycerol to

600.0mL of distilled/deionized water. Mix thorough

ly. Add remaining components, except egg mixture,

lincomycin solution, cycloheximide solution, and na

lidixic acid solution. Mix thoroughly. Bring volume

to 1.0L. Gently heat while stirring and bring to boil

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

Cool to 50°C. Aseptically add 625.0mL of egg mix

ture. Mix thoroughly. Aseptically add 1.0mL cyclo

heximide solution, 1.0mL lincomycin solution, and

1.0mL nalidixic acid solution. Distribute into sterile

screw-capped tubes. Place tubes in a slanted position.

Inspissate at 85°C (moist heat) for 45 min.

Use: For the isolation and cultivation of Mycobacteri

um tuberculosis.

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Coletsos Selective Medium