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Thermoanaeromonas Medium (DSMZ Medium 963)

[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]

Thermoanaeromonas Medium

(DSMZ Medium 963)

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

Composition per liter:

NaHCO3 ........................................................................................ 5.0g

K2HPO4....................................................................................... 0.78g

KH2PO4....................................................................................... 0.75g

NH4Cl ........................................................................................... 0.5g

Glucose ......................................................................................... 0.5g

Yeast extract.................................................................................. 0.5g

MgSO4·7H2O.............................................................................. 0.25g

NaCl.............................................................................................. 0.2g

Na3-EDTA................................................................................... 0.04g

CaCl2·2H2O ................................................................................ 0.03g

FeSO4·7H2O................................................................................ 0.01g

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

Thiosulfate solution .................................................................20.0mL

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

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

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

pH 6.5 ± 0.2 at 25°C

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.

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.

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O ................................................................... 0.25g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to

distilled/deionized water and bring volume to 10.0mL. Mix thorough

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

121°C.

Thiosulfate Solution:

Composition per 20.0mL:

Na2S2O3·5H2O ............................................................................ 1.24g

Preparation of Thiosulfate Solution: Add Na2S2O3·5H2O to dis

tilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Au

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

Preparation of Medium: Add components, except NaHCO3, vita

min solution, thiosulfate solution, and cysteine solution, to distilled/de

ionized water and bring volume to 960.0mL. Mix thoroughly. Gently

heat and bring to boiling. Boil for 3 min. Cool to 25°C while sparging

with 80% N2 + 20% CO2. Add solid NaHCO3. Adjust pH to 6.8–7.0.

Distribute to anaerobe tubes or bottles under 80% N2 + 20% CO2. Au

toclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobi

cally add, per liter of medium, 10.0mL sterile vitamin solution, 10.0mL

sterile cysteine solution, and 20.0mL sterile thiosulfate solution. Mix

thoroughly. The final pH should be 6.5.

Use: For the cultivation of Thermanaeromonas toyohensis.

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Thermoanaeromonas Medium (DSMZ Medium 963)