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Tepidanaerobacter Medium (DSMZ Medium 1051)

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

Tepidanaerobacter Medium

(DSMZ Medium 1051)

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 ................................................................................. 2.3g

NH4Cl ........................................................................................ 0.54g

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

CaCl2·2H2O ................................................................................ 0.15g

KH2PO4....................................................................................... 0.14g

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

Bicarbonate solution ................................................................10.0mL

Glucose solution ......................................................................10.0mL

Sulfide solution........................................................................10.0mL

Cysteine solution .....................................................................10.0mL

Trace elements solution .............................................................1.0mL

Vitamin solution.........................................................................2.0mL

pH 7.1 ± 0.2 at 25°C

Trace Elements Solution:

Composition per liter:

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

FeCl3·6H2O................................................................................. 1.35g

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

NiCl2·6H2O................................................................................. 0.12g

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

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

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

Na2SeO3·5H2O.......................................................................... 0.026g

CuCl2·2H2O .............................................................................. 0.025g

CoCl2·6H2O .............................................................................. 0.024g

Na2MoO4·2H2O ........................................................................ 0.024g

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

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to approximately 500.0mL of distilled/deionized water. Dissolve

by adding KOH and adjust pH to 6.5. Add remaining components.

Bring volume to 1.0L with additional distilled/deionized water. Adjust

pH to 7.0 with KOH.

Glucose Solution:

Composition per 10.0mL:

D-Glucose...................................................................................... 2.2g

Preparation of Glucose Solution: Add glucose to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with

100% N2. Filter sterilize.

Sulfide Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.3g

Preparation of Sulfide Solution: Add Na2S·9H2O to distilled/de

ionized 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.

Bicarbonate Solution:

Composition per 10.0mL:

NaHCO3 ........................................................................................ 2.5g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

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

with a gas mixture of 80% N2 + 20% CO2. Filter sterilize.

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine-HCl·2H2O ................................................................... 0.3g

Preparation of Cysteine Solution: Add L-cysteine to distilled/de

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

with 100% N2. Filter sterilize.

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 100% N2. Filter sterilize.

Preparation of Medium: Add components, except vitamin, bicar

bonate, glucose, cysteine, and sulfide solutions, to distilled/deionized

water and bring volume to 958.0mL. Mix thoroughly. Gently heat and

bring to boiling. Boil for 1 min. Cool to room temperature while sparg

ing with a gas mixture of 80% N2 + 20% CO2. Dispense into culture

vessels under an atmosphere of 80% N2 + 20% CO2. Autoclave for 15

min at 15 psi pressure–121°C. Cool to room temperature. Aspetically

add vitamin, bicarbonate, glucose, cysteine, and sulfide solutions. Mix

thoroughly. Adjust the pH to 7.0–7.2.

Use: For the cultivation of Tepidanaerobacter syntrophicus.

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Tepidanaerobacter Medium (DSMZ Medium 1051)