Mycobactosel™ L-J Medium

[所属分类:培养基配方] [发布时间:2021-6-15] [发布人:网站管理员2] [阅读次数:] [返回]

Mycobactosel™ L-J Medium

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

Composition per liter:

Potato flour .................................................................................30.0g
L- Asparagine .................................................................................3.6g
KH 2 PO 4 , anhydrous......................................................................2.5g
Sodium citrate...............................................................................0.6g
MgSO 4 ·7H 2 O..............................................................................0.24g
Homogenized whole egg ..............................................................1.0L
Malachite Green solution.........................................................20.0mL
Glycerol ...................................................................................12.0mL
Antibiotic solution ...................................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Source:  This medium is available as a prepared medium from BD Di-
agnostic Systems.
Homogenized Whole Egg:
Composition per liter:
Whole eggs ................................................................................18–24
Preparation of Whole Egg:  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. Homogenize by shaking. Filter through
four layers of sterile cheesecloth into a sterile graduated cylinder. Mea-
sure out 1.0L.
Malachite Green Solution:
Composition per 20.0mL:
Malachite Green............................................................................0.4g
Preparation of Malachite Green Solution:  Add Malachite
Green to sterile distilled/deionized water and bring volume to 20.0mL
in a sterile container. Mix thoroughly.
Antibiotic Solution:
Composition per 10.0mL:
Cycloheximide............................................................................0.64g
Nalidixic acid............................................................................0.056g
Lincomycin................................................................................3.2mg
Preparation of Antibiotic Solution: Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
Caution:  Cycloheximide is toxic. Avoid skin contact or aerosol for-
mation and inhalation.
Preparation of Medium: Add components—except whole egg,
Malachite Green solution, and antibiotic solution—to distilled/deion-
ized water and bring volume to 600.0mL. Mix thoroughly. Autoclave
for 30 min at 15 psi pressure–121°C. Cool to room temperature. Add
the homogenized whole egg, Malachite Green solution, and antibiotic
solution. Distribute into sterile tubes in 8.0mL volumes. Coagulate me-
dium in a slanted position at 85°C (moist heat) for 50 min.
Use: For the isolation and cultivation of Mycobacterium species from
clinical specimens.