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培养基配方
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Treponema denticola Medium (DSMZ Medium 909)
[所属分类:培养基配方] [发布时间:2021-9-18] [发布人:网站管理员2] [阅读次数:] [返回]
Treponema denticola Medium
(DSMZ Medium 909)
山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com
Composition per 1045.0mL:
Solution A.....................................................................................1.0L
Solution B ................................................................................45.0mL
pH 7.1 ± 0.2 at 25°C
Solution A:
Composition per liter:
Trypticase™................................................................................ 10.0g
Yeast extract.................................................................................. 2.5g
Agar .............................................................................................. 3.0g
Glucose ......................................................................................... 2.0g
L-Cysteine·HCl ............................................................................. 1.0g
Na-thioglycolate ........................................................................... 0.5g
L-Asparagine............................................................................... 0.25g
Brain heart infusion broth......................................................450.0mL
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring
to boiling. Boil for 5 min. Cool to room temperature while sparging
with 80% N2 + 20% CO2. Distribute to anaerobe tubes or bottles under
80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to room temperature.
Brain Heart Infusion Broth:
Composition per liter:
Pancreatic digest of gelatin ......................................................... 14.5g
Brain heart, solids from infusion .................................................. 6.0g
Peptic digest of animal tissue........................................................ 6.0g
NaCl .............................................................................................. 5.0g
Casein............................................................................................ 5.0g
Glucose ......................................................................................... 3.0g
Na2HPO4 ....................................................................................... 2.5g
Preparation of Brain Heart Infusion Broth: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Solution B:
Composition per 65.0mL:
NaHCO3 ........................................................................................ 2.0g
Thiamine pyrophosphate............................................................6.0mg
Volatile fatty acid solution .......................................................20.0mL
Rabbit serum ............................................................................20.0mL
Volatile Fatty Acid Solution:
Composition per 102.0mL:
KOH, 0.1N.............................................................................100.0mL
Isobutyric acid............................................................................0.5mL
D,L-2-methylbutyric acid ...........................................................0.5mL
Isovaleric acid ............................................................................0.5mL
Valeric acid ................................................................................0.5mL
Preparation of Volatile Fatty Acid Solution: Combine compo
nents and mix thoroughly.
Preparation of Solution B: Add components, except rabbit serum,
to distilled/deionized water and bring volume to 45.0mL. Mix thor
oughly. Filter sterilize. Add 20.0mL rabbit serum.
Preparation of Medium: Aseptically and anaerobically add
0.45mL solution B and 10.0mL solution A to individual tubes. Pressur
ize the tubes with H2 to 0.5 bar.
Use: For the cultivation of Treponema denticola.