Differential Agar Medium A8 for Ureaplasma urealyticum

[所属分类:培养基配方] [发布时间:2020-6-19] [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

Differential Agar Medium A8
for Ureaplasma urealyticum

培养基配方
Composition per 103.1mL:
Basal agar..............................................................80.0mL
Horse serum, unheated...................................................20.0mL
Fresh yeast extract solution.............................................1.0mL
Urea solution............................................................1.0mL
CVA enrichment...........................................................0.5mL
L- Cysteine·HCl·H2O solution...........................................0.5mL
GHL tripeptide solution..................................................0.1mL
pH 5.5 ± 0.2 at 25°C
Basal Agar:
Composition per 80.0mL:
Tryptic soy broth.........................................................2.4g
Noble agar...............................................................1.05g
Putrescine·2HCl.........................................................0.17g
CaCl2 ·2H2O............................................................0.015g
Preparation of Basal Agar: Add components to distilled/deion-
ized water and bring volume to 80.0mL. Mix thoroughly. Adjust pH to
5.5 with 2N HCl. Gently heat and bring to boiling. Autoclave for 15
min at 15 psi pressure–121°C. Cool to 50°–55°C.
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. Filter sterilize.
Urea Solution:
Composition per 30.0mL:
Urea...................................................................3.0g
Preparation of Urea Solution: Add urea to distilled/deionized wa-
ter and bring volume to 30.0mL. Mix thoroughly. Filter sterilize.
CVA Enrichment:
Composition per liter:
Glucose .............................................................100.0g
L- Cysteine·HCl·H2O.................................................25.9g
L- Glutamine..........................................................10.0g
Adenine................................................................1.0g
L- Cystine·2HCl.......................................................1.0g
Nicotinamide adenine dinucleotide ....................................0.25g
Cocarboxylase..........................................................0.1g
Guanine·HCl..........................................................0.03g
Fe(NO3)3 .............................................................0.02g
Vitamin B 12 .........................................................0.01g
p-Aminobenzoic acid..................................................0.013g
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·H2O Solution:
Composition per 50.0mL:
L -Cysteine·HCl·H2O ................................................1.0g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -cyste-
ine·HCl·H2O to distilled/deionized water and bring volume to 50.0mL.
Mix thoroughly. Filter sterilize.
GHL Tripeptide Solution:
Composition per 10.0mL:
GHL tripeptide.......................................................0.2mg
Preparation of GHL Tripeptide Solution: Add GHL tripeptide
(glycyl- L- histidyl- L- lysine acetate) to distilled/deionized water and
bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: To 80.0mL of cooled, sterile basal agar,
aseptically add 20.0mL of sterile horse serum, 1.0mL of sterile fresh
yeast extract solution, 1.0mL of sterile urea solution, 0.5mL of sterile
CVA enrichment, 0.5mL of sterile  L -cysteine·HCl·H2O solution, and
0.1mL of sterile GHL tripeptide solution. Mix thoroughly. Pour into
sterile Petri dishes in 20.0mL volumes.
Use:  For the cultivation and maintenance of Ureaplasma urealyticum.