​Avian Mycoplasma Agar

[所属分类:培养基配方] [发布时间:2019-11-15] [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

Avian Mycoplasma Agar
Composition per liter:
Agar, not inhibitory to mycoplasmas..........................................10.0g
PPLO broth without Crystal Violet........................................700.0mL
Swine or horse serum, heat inactivated
at 56°C for 30 min...........................................................150.0mL
Fresh yeast extract solution ...................................................100.0mL
Phenol Red solution.................................................................20.0mL
Glucose solution......................................................................10.0mL
Arginine solution.....................................................................10.0mL
NAD solution...........................................................................10.0mL
PPLO Broth without Crystal Violet:
Composition per 700.0mL:
Beef heart, infusion from..........................................................175.0g
Peptone .........................................................................................7.0g
NaCl..............................................................................................3.5g
Source: PPLO broth without Crystal Violet is available as a premixed
powder from BD Diagnostic Systems.
Preparation of PPLO Broth without Crystal Violet:  Add
components to distilled/deionized water and bring volume to 700.0mL.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Beef
heart for infusion may be substituted; 100.0g of beef heart for infusion
is equivalent to 500.0g of fresh heart tissue.
Fresh Yeast Extract Solution:
Composition per 100.0mL:
Baker’s yeast, live, pressed, starch-free......................................25.0g
Preparation of Fresh Yeast Extract Solution : Add the live Bak-
er’s yeast to 100.0mL of distilled/deionized water. Autoclave for 90
min at 15 psi pressure–121°C. Allow to stand. Remove supernatant so-
lution. Adjust pH to 6.6–6.8.
Phenol Red Solution:
Composition per 20.0mL:
Phenol Red..................................................................................0.02g
Preparation of Phenol Red Solution:  Add Phenol Red to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Filter sterilize.
Glucose Solution:
Composition per 10.0mL:
Glucose .........................................................................................1.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize.
Arginine Solution:
Composition per 10.0mL:
Arginine ........................................................................................1.0g
Preparation of Arginine Solution:  Add arginine to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
NAD Solution:
Composition per 10.0mL:
NAD..............................................................................................0.1g
Preparation of NAD Solution:  Add NAD to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add 10.0g of agar to 700.0mL of PPLO
broth without Crystal Violet. Gently heat to boiling with frequent mixing.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Warm
other components to 50°–55°C using a water bath. Aseptically combine
all components. Mix thoroughly. Pour into sterile Petri dishes or sterile
tubes.
Use: For the cultivation and maintenance of Mycoplasma species.