PPLO Agar with Additives for Mycoplasma

[所属分类:培养基配方] [发布时间:2021-7-13] [发布人:网站管理员2] [阅读次数:] [返回]

PPLO Agar with Additives for Mycoplasma

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

Composition per 1010.0mL:

Agar ............................................................................................15.0g
Arginine ......................................................................................1.74g
Glutamine....................................................................................1.46g
Phenol Red..................................................................................0.02g
PPLO broth............................................................................700.0mL
Horse serum, not inactivated .................................................200.0mL
Yeast extract solution, fresh...................................................100.0mL
Vitamins in Eagle’s medium, 100X.........................................10.0mL
pH 7.1 ± 0.2 at 25°C
PPLO Broth:
Composition per liter:
Beef heart, infusion from............................................................50.0g
Peptone........................................................................................10.0g
NaCl..............................................................................................5.0g
Preparation of PPLO Broth: Add components to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly. Gently heat and
bring to boiling.
Yeast Extract Solution:
Composition per 300.0mL:
Baker’s yeast, live, pressed, starch-free......................................75.0g
Preparation of Yeast Extract Solution : Add the live Baker’s
yeast to 300.0mL of distilled/deionized water. Autoclave for 90 min at
15 psi pressure–121°C. Allow to stand. Remove supernatant solution.
Adjust pH to 6.6–6.8. Filter sterilize.
Vitamins in Eagle’s Medium, 100X:
Composition per liter:
Inositol.......................................................................................2.0mg
Calcium pantothenate ................................................................1.0mg
Choline chloride.........................................................................1.0mg
Folic acid ...................................................................................1.0mg
Nicotinamide..............................................................................1.0mg
Pyridoxal....................................................................................1.0mg
Thiamine·HCl............................................................................1.0mg
Riboflavin..................................................................................0.1mg
Preparation of Vitamins in Eagle’s Medium, 100X: Add com-
ponents to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Filter sterilize.
Preparation of Medium: Add components—except fresh yeast ex-
tract solution, horse serum, and vitamins in Eagle’s medium, 100X—
to distilled/deionized water and bring volume to 690.0mL. Mix thor-
oughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 45°–50°C. Aseptically add sterile fresh yeast
extract solution, horse serum, and vitamins in Eagle’s medium, 100X.
Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile
tubes.
Use:  For the cultivation and maintenance of Mycoplasma arginini and
Spiroplasma apis.