A 7 Agar (Shepard’s Differential Agar)

[所属分类:培养基配方] [发布时间:2019-9-16] [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

A 7 Agar
(Shepard’s Differential Agar)
Composition per 205.7mL:
Agar base ...............................................................................160.0mL
Supplement solution ................................................................45.7mL
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
MnSO 4 ·H 2 O................................................................................0.15g
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 45.72mL:
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............................................................................0.22mL
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. Filter sterilize.
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
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·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, ultrapure..............................................................................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 160.0mL of cooled,
sterile agar base and 45.9mL of sterile supplement solution. Mix thor-
oughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use: For the cultivation and differentiation of Ureaplasma urealyti-
cum from urine based on its ability to produce ammonia from urea.
Bacteria that produce ammonia appear as golden to dark brown colo-
nies. Also used for the cultivation of other Ureaplasma species.