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Cefsulodin Irgasan Novobiocin Agar (CIN Agar) (Yersinia Selective Agar) (BAM M35)

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

Cefsulodin Irgasan Novobiocin Agar

(CIN Agar)

(Yersinia Selective Agar)

(BAM M35)

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

Composition per 1008.0mL:

Basal medium ............................................. 757.0mL

Deoxycholate solution ................................ 200.0mL

Cefsulodin solution....................................... 10.0mL

Novobiocin solution...................................... 10.0mL

Crystal Violet solution .................................. 10.0mL

Strontium chloride solution .......................... 10.0mL

Neutral Red solution..................................... 10.0mL

NaOH, 5N ....................................................... 1.0mL

Irgasan solution............................................... 1.0mL

pH 7.4 ± 0.2 at 25°C

Basal Medium:

Composition per 757.0mL:

Mannitol............................................................20.0g

Special peptone.................................................20.0g

Agar ..................................................................12.0g

Sodium pyruvate.................................................2.0g

Yeast extract........................................................2.0g

NaCl....................................................................1.0g

Magnesium sulfate solution............................ 1.0mL

Preparation of Basal Medium: Add compo

nents to distilled/deionized water and bring volume

to 757.0mL. Mix thoroughly. Gently heat and bring

to boiling with stirring. Cool to about 80°C by plac

ing in a 50°C water bath for about 10 min.

Magnesium Sulfate Solution:

Composition per 10mL:

MgSO4·7H2O......................................................0.1g

Preparation of Magnesium Sulfate Solution:

Add MgSO4·7H2O to distilled/deionized water and

bring volume to 10.0mL. Mix thoroughly.

Irgasan Solution:

Composition per 10mL:

Irgasan (triclosan...............................................0.04g

Preparation of Irgasan Solution: Add irgasan

to 95% ethanol and bring volume to 10.0mL. Mix

thoroughly. Can be stored for 4 weeks at –20°C.

Deoxycholate Solution:

Composition per 200.0mL:

Na-deoxycholate .................................................0.5g

Preparation of Deoxycholate Solution: Add

deoxycholate to distilled/deionized water and bring

volume to 200.0mL. Mix thoroughly. Gently heat

and bring to boiling with stirring. Cool to 50–55°C.

Neutral Red Solution:

Composition per 10.0mL:

Neutral Red....................................................30.0mg

Preparation of Neutral Red Solution: Add

Neutral Red to 10.0mL of distilled/deionized water.

Mix thoroughly. Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to 25°C.

Crystal Violet Solution:

Composition per 10.0mL:

Crystal Violet ...................................................1.0mg

Preparation of Crystal Violet Solution: Add

Crystal Violet to 10.0mL of distilled/deionized water.

Mix thoroughly. Autoclave for 15 min at 15 psi pres

sure–121°C. Cool to 25°C.

Cefsulodin Solution:

Composition per 10.0mL:

Cefsulodin......................................................15.0mg

Preparation of Cefsulodin Solution: Add cefsu

lodin to 10.0mL of distilled/deionized water. Mix thor

oughly. Filter sterilize.

Novobiocin Solution:

Composition per 10.0mL:

Novobiocin.......................................................2.5mg

Preparation of Novobiocin Solution: Add novo

biocin to 10.0mL of distilled/deionized water. Mix

thoroughly. Filter sterilize.

Strontium Chloride Solution:

Composition per 10.0mL:

SrCl2·6H2O .........................................................1.0g

Preparation of Strontium Chloride: Add stron

tium chloride to 10.0mL of distilled/deionized water.

Mix thoroughly. Filter sterilize.

Preparation of Medium: Add 1.0mL irgasan so

lution to 757.0mL basal medium. Mix thoroughly.

Cool to 50–55°C. Add 200.0mL desoxychlolate solu

tion. Mix thoroughly. Solution should remain clear.

Aseptically add 1.0mL 5N NaOH, 10.0mL Neutral

Red solution, 10.0mL Crystal Violet solution,

10.0mL cefsulodin solution, and 10.0mL novobiocin

solution. Mix thoroughly. Slowly add 10.0mL stron

tium chloride solution while continuously stirring.

Adjust pH to 7.4 with 5N NaOH. Pour into sterile

Petri dishes or distribute into sterile tubes.

Use: For the selective isolation and differentiation of

Yersinia enterocolitica based on mannitol fermenta

tion. Yersinia enterocolitica appears as “bull’s eye”

colonies with deep red centers surrounded by a trans

parent periphery.

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Cefsulodin Irgasan Novobiocin Agar (CIN Agar) (Yersinia Selective Agar) (BAM M35)