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Treponema saccharophilum 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:

Pancreatic digest of casein............................................................ 2.0g

Yeast extract.................................................................................. 2.0g

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

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

Salt solution A .......................................................................200.0mL

Salt solution B........................................................................200.0mL

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

Glucose solution ......................................................................20.0mL

iso-Butyric acid..........................................................................0.4mL

n-Butyric acid ............................................................................0.4mL

DL-2-Methylbutyric acid ............................................................0.2mL

iso-Valeric acid ..........................................................................0.2mL

n-Valeric acid .............................................................................0.2mL

pH 6.7–7.0 at 25°C

Salt Solution A:

Composition per liter:

MgSO4·7H2O .............................................................................. 0.96g

CaCl2·2H2O................................................................................. 0.59g

Preparation of Salt Solution A: Add components to distilled/de

ionized water and bring volume to 1.0L. Mix thoroughly.

Salt Solution B:

Composition per liter:

K2HPO4...................................................................................... .2.25g

KH2PO4....................................................................................... 2.25g

NaCl .............................................................................................. 4.5g

(NH4)2SO4..................................................................................... 4.5g

Preparation of Salt Solution B: Add components to distilled/de

ionized water and bring volume to 1.0L. Mix thoroughly.

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 steril

ize. Sparge with 100% CO2.

Preparation of Medium: Prepare and dispense medium under

100% CO2. Add components, except NaHCO3 solution and glucose so

lution, to distilled/deionized water and bring volume to 880.0mL. Mix

thoroughly. Adjust pH to 7.0 with KOH. Gently heat and bring to boil

ing. Continue boiling for 5 min. Cool to room temperature while sparg

ing with 100% CO2. Anaerobically distribute 8.8mL into anaerobic

culture tubes. Autoclave for 15 min at 15 psi pressure–121°C. Using a

syringe, add 1.0mL of sterile NaHCO3 solution and 0.2 mL of sterile

glucose solution to each tube. Check that final pH is 6.7–7.0.

Use: For the cultivation and maintenance of Treponema saccharophi

lum.

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Treponema saccharophilum Medium