AH5 Medium

[所属分类:培养基配方] [发布时间:2019-9-23] [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

AH5 Medium
Composition per 205.9mL:
Agar base...............................................................................160.0mL
Supplement solution ................................................................45.9mL
pH 6.0 ± 0.2 at 25°C
Agar Base:
Composition per 165.0mL:
Pancreatic digest of casein..........................................................2.72g
Agar..............................................................................................2.1g
NaCl..............................................................................................0.8g
Papaic digest of soybean meal....................................................0.48g
K 2 HPO 4 .........................................................................................0.4g
Glucose.........................................................................................0.4g
Preparation of Agar Base:  Add components, except agar, to dis-
tilled/deionized water and bring volume to 165.0mL. Adjust pH to 5.5.
Add agar. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 45°–50°C.
Supplement Solution:
Composition per 45.9mL:
Horse serum, unheated.............................................................40.0mL
Fresh yeast extract solution .......................................................2.0mL
Penicillin solution......................................................................2.0mL
CVA enrichment ........................................................................1.0mL
L -Cysteine·HCl·H 2 O solution....................................................0.5mL
Urea solution..............................................................................0.4mL
Preparation of Supplement Solution:  Aseptically combine com-
ponents. Mix thoroughly.
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.
Penicillin Solution:
Composition per 10.0mL:
Penicillin G.......................................................................1,000,000U
Preparation of Penicillin Solution: Add penicillin to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
CVA Enrichment:
Composition per liter:
Glucose .....................................................................................100.0g
L- Cysteine·HCl·H 2 O....................................................................25.9g
L- Glutamine.................................................................................10.0g
L- Cystine·2HCl..............................................................................1.0g
Adenine.........................................................................................1.0g
Nicotinamide adenine dinucleotide ............................................0.25g
Cocarboxylase...............................................................................0.1g
Guanine·HCl...............................................................................0.03g
Fe(NO 3 ) 3 .....................................................................................0.02g
p-Aminobenzoic acid................................................................0.013g
Vitamin B 12 .................................................................................0.01g
Thiamine·HCl ............................................................................3.0mg
Preparation of CVA Enrichment: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L- Cysteine·HCl·H 2 O......................................................................0.4g
Preparation of  L -Cysteine·HCl·H 2 O Solution: Add  L-
cysteine·HCl·H 2 O solution to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.
Urea Solution:
Composition per 10.0mL:
Urea...............................................................................................1.0g
Preparation of Urea Solution: Add urea to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium:  Aseptically combine cooled, sterile com-
ponents. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation of Ureaplasma urealyticum from urine and
exudates and for the cultivation of other Ureaplasma species.