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Thermodesulfobacterium 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 liter:

Na2SO4.......................................................................................... 3.0g

Na2HPO4·12H2O........................................................................... 2.0g

NH4Cl ........................................................................................... 1.0g

KH2PO4......................................................................................... 0.3g

MgCl2·6H2O ................................................................................. 0.2g

FeSO4·7H2O............................................................................... 1.5mg

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

Sodium lactate solution............................................................20.0mL

Trace elements solution ...........................................................10.0mL

Yeast extract solution...............................................................10.0mL

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

Vitamin solution.........................................................................5.0mL

pH 6.8 ± 0.2 at 25°C

Sodium Lactate Solution:

Composition per 20.0mL:

Sodium lactate .............................................................................. 4.0g

Preparation of Sodium Lactate Solution: Add sodium lactate to

distilled/deionized water and bring volume to 20.0mL. Mix thorough

ly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

Trace Elements Solution:

Composition per liter:

Nitrilotriacetic acid ..................................................................... 12.8g

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

FeCl·4H2O .................................................................................... 0.20

CoCl2·6H2O................................................................................ 0.17g

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

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

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

CuCl2 .......................................................................................... 0.02g

H3BO3......................................................................................... 0.01g

Na2MoO4·2H2O.......................................................................... 0.01g

NiCl2·6H2O............................................................................... 0.026g

Na2SeO3·5H2O............................................................................ 0.02g

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with

KOH. Add remaining components. Add distilled/deionized water to

1.0L.

Yeast Extract Solution:

Composition per 10.0mL:

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

Preparation of Yeast Extract Solution: Add yeast extract to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

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

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. Mix thoroughly.

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

Prior to use, neutralize solution by dropwise addition of sterile 1N HCl.

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ........................................................................10.0mg

Calcium DL-pantothenate...........................................................5.0mg

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

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

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

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

Thiamine·HCl ............................................................................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.

Preparation of Medium: Prepare and dispense medium under

100% N2. Add components, except sodium lactate solution, yeast ex

tract solution, and Na2S·9H2O solution, to distilled/deionized water

and bring volume to 960.0mL. Mix thoroughly. Sparge with 100% N2.

Anaerobically distribute into tubes or bottles. Autoclave for 15 min at

15 psi pressure–121°C. Aseptically and anaerobically add to 1.0L of

medium 20.0mL of sterile sodium lactate solution, 10.0mL of sterile

yeast extract solution, and 10.0mL of sterile Na2S·9H2O solution. Mix

thoroughly.

Use: For the cultivation and maintenance of Thermodesulfobacterium

commune.

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Thermodesulfobacterium Medium