Coletsos Selective Medium

[所属分类:培养基配方] [发布时间:2020-5-21] [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

Coletsos Selective Medium

培养基配方
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
Egg mixture solution.......................................................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 thorough-
ly. 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 thoroughly. Bring volume to 100.0mL with
distilled/deionized water. Filter sterilize.
Caution:  Cycloheximide is toxic. Avoid skin contact or aerosol for-
mation and inhalation.
Lincomycin Solution:
Composition per 100.0mL:
Lincomycin.................................................................0.5g
Preparation of Lioncomycin Solution: Add lincomycin to dis-
tilled/deionized water and bring volume 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 ster-
ile graduated cylinder. Bring volume to 1.0L with distilled/deionized
water.
Preparation of Medium: Add glycerol to 600.0mL of distilled/de-
ionized water. Mix thoroughly. Add remaining components, except egg
mixture, lincomycin solution, cycloheximide solution, and nalidixic
acid solution. Mix thoroughly. Bring volume to 1.0L. Gently heat while
stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 50°C. Aseptically add 625.0mL of egg mixture. Mix
thoroughly. Aseptically add 1.0mL cycloheximide solution, 1.0mL lin-
comycin 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 Mycobacterium tuberculosis.