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

[所属分类:培养基配方] [发布时间:2020-4-13] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

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

培养基配方
Composition per 1008.0mL:
Basal medium ............................................757.0mL
Desoxycholate 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 components to distilled/de-
ionized water and bring volume to 757.0mL. Mix thoroughly. Gently
heat and bring to boiling with stirring. Cool to about 80°C by placing
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 thor-
oughly.
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.
Desoxycholate Solution:
Composition per 200.0mL:
Na-desoxycholate .............................................0.5g
Preparation of Desoxycholate Solution:  Add desoxycholate to
distilled/deionized water and bring volume to 200.0mL. Mix thorough-
ly. 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 pressure–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 pressure–121°C. Cool to 25°C.
Cefsulodin Solution:
Composition per 10.0mL :
Cefsulodin..................................................15.0mg
Preparation of Cefsulodin Solution: Add cefsulodin to 10.0mL of
distilled/deionized water. Mix thoroughly. Filter sterilize.
Novobiocin Solution:
Composition per 10.0mL :
Novobiocin...................................................2.5mg
Preparation of Novobiocin Solution: Add novobiocin 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 strontium chloride to
10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add 1.0mL irgasan solution to 757.0mL
basal medium. Mix thoroughly. Cool to 50–55°C. Add 200.0mL des-
oxychlolate solution. 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
strontium chloride solution while continuously stirring. Adjust pH to
7.4 with 5N NaOH. Pour into sterile Petri dishes or distribute into ster-
ile tubes.
Use : For the selective isolation and differentiation of Yersinia entero-
colitica based on mannitol fermentation. Yersinia enterocolitica
appears as “bull’s eye” colonies with deep red centers surrounded by a
transparent periphery.