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Treponema succinifaciens Medium (DSMZ Medium 275)

[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 succinifaciens Medium

(DSMZ Medium 275)

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

Composition per liter:

Solution A ..............................................................................875.0mL

Solution B ................................................................................50.0mL

Solution D ................................................................................50.0mL

Solution C ................................................................................25.0mL

pH 7.0 ± 0.2 at 25°C

Solution A:

Composition per 875.0mL:

NaCl .............................................................................................. 1.0g

K2HPO4......................................................................................... 0.5g

Cysteine-HCl·H2O ........................................................................ 0.5g

KH2PO4......................................................................................... 0.5g

Yeast extract.................................................................................. 0.5g

Peptone ......................................................................................... 0.5g

(NH4)2SO4 .................................................................................... 0.5g

CaCl2·2H2O .................................................................................. 0.1g

MgSO4·7H2O................................................................................ 0.1g

Resazurin .................................................................................. 0.001g

Rumen fluid, clarified............................................................300.0mL

Preparation of Solution A: Add components to 575.0mL of dis

tilled/deionized water. Mix thoroughly. Adjust pH to 6.2–6.3 with 4N

HCl. Heat to boiling point. Cool to room temperature under 100% N2.

Autoclave for 15 min at 15 psi pressure–121°C.

Solution B:

Composition per 100.0mL:

Glucose ....................................................................................... 20.0g

Preparation of Solution B: Add glucose to 100.0mL of distilled/de

ionized water. Mix thoroughly. Sparge with 100% N2. Autoclave for 15

min at 15 psi pressure–121°C.

Solution C:

Composition per 100.0mL:

KH2PO4....................................................................................... 0.45g

Na2HPO4·2H2O........................................................................... 0.58g

Preparation of Solution C: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.4.

Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Solution D:

Composition per 100.0mL:

NaHCO3........................................................................................ 5.0g

Preparation of Solution D: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80%

N2 + 20% CO2. Filter sterilize.

Preparation of Medium: Distribute solution A under 100% N2 into

anaerobic tubes. Autoclave for 20 min at 15 psi pressure–121°C. Cool

to 25°C. Aseptically and anaerobically add appropriate amounts of so

lutions B, C, and D to achieve final concentrations.

Use: For the cultivation of Treponema succinifaciens.

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Treponema succinifaciens Medium (DSMZ Medium 275)