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Thermoanaerobacter subterraneus Medium (DSMZ Medium 899)

[所属分类:培养基配方] [发布时间:2021-9-6] [发布人:网站管理员2] [阅读次数:] [返回]

Thermoanaerobacter subterraneus Medium

(DSMZ Medium 899)

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

Composition per liter:

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

MgCl2·6H2O.................................................................................. 1.0g

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

NaCl .............................................................................................. 0.6g

Cysteine-HCl·H2O ........................................................................ 0.5g

K2HPO4......................................................................................... 0.3g

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

KCl................................................................................................ 0.2g

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

Resazurin ...................................................................................0.5mg

D-Glucose solution...................................................................30.0mL

NaHCO3 solution .....................................................................20.0mL

Trace mineral solution .............................................................10.0mL

Na2S2O3 solution......................................................................10.0mL

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

pH 7.0 ± 0.2 at 25°C

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................. 0.45g

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

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

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

to room temperature.

NaHCO3 Solution:

Composition per 20.0mL:

NaHCO3........................................................................................ 4.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Filter sterilize.

Glucose Solution:

Composition per 30.0mL:

D-Glucose...................................................................................... 4.0g

Preparation of Glucose Solution: Add D-glucose to distilled/de

ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge

with 100% N2. Filter sterilize.

Na2S2O3 Solution:

Composition per 10.0mL:

Na2S2O3·5H2O .............................................................................. 2.5g

Preparation of Na2S2O3 Solution: Add Na2S2O3·5H2O to distilled/

deionized water and bring volume to 10.0mL. Mix thoroughly. Auto

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

room temperature.

Trace Elements Solution:

Composition per liter:

MgSO4·7H2O................................................................................ 3.0g

Nitrilotriacetic acid ....................................................................... 1.5g

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

MnSO4·2H2O................................................................................ 0.5g

CoSO4·7H2O............................................................................... 0.18g

ZnSO4·7H2O ............................................................................... 0.18g

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

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

NiCl2·6H2O............................................................................... 0.025g

KAl(SO4)2·12H2O....................................................................... 0.02g

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

Na2MoO4·4H2O .......................................................................... 0.01g

CuSO4·5H2O............................................................................... 0.01g

Na2SeO3·5H2O........................................................................... 0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH

to 6.5 with KOH. Add remaining components. Add distilled/deionized

water to 1.0L. Mix thoroughly.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3 solu

tion, glucose solution, Na2S·9H2O solution, and Na2S2O3 solution, to

distilled/deionized water and bring volume to 930.0mL. Mix thoroughly.

Sparge with 80% N2 + 20% CO2. Distribute into sterile tubes or bottles.

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

bically per 1.0L of medium add 20.0mL NaHCO3 solution, 30.0mL glu

cose solution, 10.0mL Na2S·9H2O solution, and 10.0mL Na2S2O3

solution. Mix thoroughly. The final pH should be 7.0.

Use: For the cultivation of Thermoanaerobacter subterraneus.

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Thermoanaerobacter subterraneus Medium (DSMZ Medium 899)