培养基配方

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Treponema bryantii Medium

[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]

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

Composition per liter:

L-Cysteine·HCl ............................................................................. 1.0g

NaCl.............................................................................................. 0.9g

(NH4)2SO4 .................................................................................... 0.9g

K2HPO4....................................................................................... 0.45g

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

MgSO4·7H2O.............................................................................. 0.18g

CaCl2·2H2O ................................................................................ 0.12g

Resazurin ................................................................................... 1.0mg

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

NaHCO3 solution ...................................................................100.0mL

Glucose solution (10% w/v) ....................................................20.0mL

pH 7.0 ± 0.2 at 25°C

NaHCO3 Solution:

Composition per 100.0mL:

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

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 100.0mL. Mix thoroughly. Filter

sterilize. Sparge with 100% CO2.

Glucose Solution:

Composition per 20.0mL:

Glucose ......................................................................................... 2.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

ized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize.

Sparge with 100% CO2.

Preparation of Medium: Prepare and dispense medium under

100% CO2. Add components, except rumen fluid, NaHCO3 solution,

and glucose solution, to distilled/deionized water and bring volume to

580.0mL. Mix thoroughly. Adjust pH to 7.0 with KOH. Sparge with

100% CO2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

room temperature. Aseptically add 300.0mL of sterile rumen fluid,

100.0mL of sterile NaHCO3 solution, and 20.0mL of sterile glucose so

lution. Mix thoroughly. Aseptically distribute into sterile tubes or

flasks.

Use: For the cultivation and maintenance of Treponema bryantii.