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Thermobacteroides leptospartum Medium

[所属分类:培养基配方] [发布时间:2021-9-7] [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 1168.1mL:

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

Trypticase™.................................................................................. 2.0g

NaOH solution ..............................................................................1.0L

Glucose solution ....................................................................113.0mL

Na2S solution ...........................................................................22.6mL

Solution A ................................................................................10.0mL

Mineral salts solution...............................................................10.0mL

Solution B ..................................................................................2.0mL

Resazurin solution......................................................................0.5mL

NaOH Solution:

Composition per liter:

NaOH ........................................................................................... 4.0g

Preparation of NaOH Solution: Add NaOH to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly.

Glucose Solution:

Composition per 100.0mL:

D-Glucose...................................................................................... 5.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

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

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

Na2S Solution:

Na2S ............................................................................................. 2.5g

Preparation of Na2S Solution: Gently heat 100.0mL of distilled/

deionized water to 100°C. Boil for 5 min. Sparge with 100% N2 for 15

min. Add the Na2S. Mix thoroughly. Sparge with 100% N2 for 10 min.

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

Solution A:

Composition per liter:

NH4Cl ....................................................................................... 100.0g

MgCl2·H2O .............................................................................. 100.0g

CaCl2·2H2O ............................................................................... 40.0g

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 4 with

HCl.

Mineral Salts Solution:

Composition per liter:

EDTA·2H2O.................................................................................. 0.5g

CoCl2·H2O .................................................................................. 0.15g

MnCl2·4H2O ................................................................................ 0.1g

FeSO4·7H2O.................................................................................. 0.1g

ZnCl2............................................................................................ 0.1g

AlCl3·H2O............................................................................... 40.0mg

Na2WO4·2H2O........................................................................ 30.0mg

CuCl2·2H2O ............................................................................ 20.0mg

NiSO4·H2O.............................................................................. 20.0mg

H2SeO3.................................................................................... 10.0mg

H3BO4 ...................................................................................... 10.0mg

NaMoO4·2H2O........................................................................ 10.0mg

Preparation of Mineral Salts Solution: Add components to dis

tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Ad

just pH to 3 with HCl.

Solution B:

Composition per liter:

K2HPO4·3H2O ......................................................................... 200.0g

Preparation of Solution B: Add K2HPO4·3H2O to distilled/deion

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

Resazurin Solution:

Composition per 100.0mL:

Resazurin ...................................................................................... 0.2g

Preparation of Resazurin Solution: Add resazurin to distilled/

deionized water and bring volume to 100.0mL. Mix thoroughly.

Preparation of Medium: Sparge 1.0L of NaOH solution with

100% CO2 for 30 min. Add 2.0g of yeast extract and 2.0g of Trypti

case™. Mix thoroughly. Add 10.0mL of solution A, 2.0mL of solution

B, 0.5mL of resazurin solution, and 10.0mL of mineral salts solution

with pipets which have been flushed a few times with 100% N2. Mix

thoroughly. Anaerobically distribute 9.0mL volumes into anaerobic

tubes fitted with butyl rubber stoppers. Autoclave for 15 min at 15 psi

pressure–121°C. One hour prior to inoculation, add 1.0mL of sterile

glucose solution and 0.2mL of sterile Na2S solution to each 9.0mL of

medium.

Use: For the cultivation of Thermobacteroides leptospartum.

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Thermobacteroides leptospartum Medium