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Thermosphaera Medium (DSMZ Medium 817)

[所属分类:培养基配方] [发布时间:2021-9-8] [发布人:网站管理员2] [阅读次数:] [返回]

Thermosphaera Medium

(DSMZ Medium 817)

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

Composition per liter:

MgCl2·6H2O ................................................................................. 2.2g

Yeast extract.................................................................................. 1.0g

Peptone ......................................................................................... 1.0g

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

KCl........................................................................................... 17.0mg

NH4Cl ...................................................................................... 12.5mg

CaCl2·2H2O ............................................................................... 7.0mg

K2HPO4·3H2O ........................................................................... 7.0mg

Resazurin ................................................................................... 0.4mg

FeCl3 ........................................................................................ 0.05mg

Vitamin solution.......................................................................10.0mL

Na2S·9H2O solution.................................................................10.0mL

NaHCO3 solution .....................................................................10.0mL

pH 6.5 ± 0.2 at 25°C

NaHCO3 Solution:

Composition per 10.0mL:

NaHCO3........................................................................................ 1.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge

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

25°C. Must be prepared freshly.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.5g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

tilled/deionized water and bring volume to 10.0mL. Sparge with N2.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an

aerobically.

Vitamin Solution:

Composition per liter:

Pyridoxine-HCl........................................................................ 10.0mg

Thiamine-HCl·2H2O.................................................................. 5.0mg

Riboflavin .................................................................................. 5.0mg

Nicotinic acid............................................................................. 5.0mg

D-Ca-pantothenate ..................................................................... 5.0mg

p-Aminobenzoic acid................................................................. 5.0mg

Lipoic acid ................................................................................. 5.0mg

Biotin ......................................................................................... 2.0mg

Folic acid ................................................................................... 2.0mg

Vitamin B12 ................................................................................ 0.1mg

Preparation of Vitamin Solution: Add components to distilled/

deionized water and bring volume to 1.0L. Mix thoroughly. Sparge

with 80% H2 + 20% CO2. Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2. Add components, except Na2S·9H2O solution, vitamin

solution, and NaHCO3 solution, to distilled/deionized water and bring

volume to 970.0mL. Mix thoroughly. Flush medium with 80% N2 +

20% CO2. for 5 min. Adjust medium pH to 6.53. Autoclave for 15 min

at 15 psi pressure–121°C. Cool to 25°C. Aseptically and anaerobically

add 10.0mL sterile Na2S·9H2O solution, 10.0mL sterile vitamin solu

tion, and 10.0mL sterile NaHCO3 solution. Mix thoroughly. Aseptical

ly and anaerobically distribute into sterile thick-walled tubes or thick

walled bottles. Pressurize with 2 bar atmosphere of 80% N2 + 20%

CO2.

Use: For the cultivation of Thermosphaera aggregans.

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Thermosphaera Medium (DSMZ Medium 817)